CN100417936C - Micro-device with magnetization axis and its method of use and application - Google Patents

Abstract

The invention discloses a micro-device with a magnetization axis and its use and application. The micro-device consists of a) a magnetizable base material and b) an optically identifiable coding pattern fabricated on the base material. . The microdevice also has a magnetization axis. The invention also provides a method and a system for using the above-mentioned micro-device to separate, detect, analyze, manipulate and synthesize compound libraries for entity molecules. The invention can be widely used in the fields of separation, detection, manipulation and compound library synthesis of entity molecules/molecules.

Description

Micro-device with magnetization axis and its method of use and application

technical field

The present invention relates to a microdevice with a magnetization axis and its preparation and application method and application, in particular to a microdevice with a magnetization axis and its use method and its composition in a system constituting a microdevice array and a synthetic compound library application.

Background technique

In the current diagnostic and research fields, especially where gene information needs to be acquired and analyzed, high-density, high-throughput biological or biochemical analysis has become a necessary research tool. These analytical methods often need to be carried out on solid phase media. Common examples of quantitative analysis performed on solid-phase media include the detection of antigens by enzyme-linked immunosorbent assay (ELISA), or the determination of mRNA expression levels by hybridization. Although the above-mentioned solid phase media are usually in the form of spherical beads or planar arrays, in fact, these solid phase media can take any appropriate form, not limited to the above two.

A planar solid phase medium, such as an array based on a glass slide or a chip, can immobilize capture molecules, such as antibodies, cDNA or other known capture molecules, at specific positions. The surface of the solid medium can be easily washed to remove unbound material. The analyte mixture is added to the solid phase medium for reaction, and the analyte can be captured on the surface of the medium and detected by a common label, such as a fluorescent dye. The identification of the analyte can be realized by the position information of the capture molecule that captures the analyte on the medium. Since the array has a flat surface, compared with the bead detection device, the array detection device is easier to design, and the cost is relatively low. One of the difficulties of planar arrays is to accurately place the capture molecules at specific positions on the plane. In order to solve this problem, manipulator spotting (Schena et al. Quantitative monitoring of gene expression patterns with a complementary DNA microarray. Science, 270: 467-470 (1995)), photolithography (Fodor et al. Light-directed, spatially addressable parallel chemical synthesis. Science, 251: 767-773 (1991)) and inkjet technology (Blanchard et al. High-density oligonucleotide arrays. Biosensors Bioelectronics, 6/7: 687-690 (1996)). But these methods have certain limitations. In order to fabricate high-density arrays (over 1000 dots/cm ² ), expensive equipment is required. Moreover, once the array is fabricated, the arrangement of the samples cannot be changed, such as replacing the capture cDNA in the array with a cDNA. If the arrangement of samples is to be changed, the array needs to be remade, which greatly increases the cost. Molecules immobilized on a planar medium react less efficiently than molecules freely distributed in solution.

Using the surface of tiny particles as a solid medium can overcome the above-mentioned problems. Small particles often choose spherical beads because they have a uniform geometric shape and are less likely to interact with each other. The problem with using microparticles is that microparticles are difficult to identify individually because the mixture of beads does not have discrete, specific positional information like the probes on the array. To address this issue, several methods have emerged to encode beads to facilitate individual identification of the beads. Some companies, such as Luminex, make the beads photorecognizable by incorporating a mixture of different fluorescent dyes into the beads. Similar to this method, some researchers have incorporated other different optically identifiable substances (such as quantum dots) in the beads so that the beads can be identified by light (Han et al. Quantum-dot-tagged microbeads for multiplexed optical coding of biomolecules. Nature Biotechnology, 19:631-635 (2001)). Quantum dots, this kind of nanoscale microparticles can also be used for the detection of beads. The special optical properties of quantum dots are related to the composition and size of quantum dots. Quantum dots of different sizes and compositions can be produced according to specific needs. Quantum dots can absorb light and immediately emit light at another frequency. The most important property of quantum dots is that as long as the size of the quantum dots is appropriately changed, the quantum dots can absorb or emit light of a specific frequency. Genicon Technologies (their Resonance Light Scattering "RLS" particles are nanoscale in size and have "resonance light scattering" properties) also develop micro- and nano-scale beads with photorecognizable properties. However, it is difficult to produce more than 1,000 kinds of beads with different coding marks no matter which method is used.

Solid media in the form of beads are also commonly used in combinatorial chemistry. In the synthesis process of a bead/a compound (also known as the separation and mixing process) (Lam et al. The "one-bead-one-compound" combinatorial library method. Chem. Rev., 97: 411- 448 (1997)), which can generate large compound libraries containing more than 10 ⁸ different molecules. The recognition of the beads is achieved by the complexes on the beads. Beads in the same group are labeled with the same compound "tea bag" to distinguish different kinds of beads. Recently, IRORI has expanded the "tea bag" marker into a small cavity with an electromagnetic wave emitter or a small cavity with an optically encoded surface. However, the compound library constructed using this technology only contains about 10,000 different compounds, and a single small cavity occupies a volume of about 0.25ml, and this technology is not suitable for high-throughput screening. PharmaSeq also uses devices with electromagnetic wave emitters. The dimensions of these devices are 250 microns by 250 microns by 100 microns. Larger libraries of compounds can be directly synthesized on a planar surface, forming a planar array, by means of photochemical lithography, such as the technique used by Affymetrix. However, this method is greatly limited by the cost of synthesis and the efficiency of multi-step synthesis in the photochemical synthesis process (McGall et al.The efficiency of light-directed synthesis of DNA arrays on glass substrates.J.Am.Chem.Soc., 119:5081-5090 (1997)), generally only applicable to the synthesis of oligonucleotides. In the past 30 years, scientists have obtained a large number of different chemical reagent-sensitive protecting groups that can be used for the synthesis of compounds on the surface of beads, but the types of photosensitive protecting groups that can be used for compound synthesis are very limited. Recently, SmartBeads Technologies introduced microfabricated barcoded particles (100 micron x 10 micron x 1 micron in size) that can be decoded and identified using a flow reader. Such microfabricated particles can easily be marked with an infinite variety of codes. The difficulty of this method lies in how to easily analyze the mixture of these encoded particles. Compared with the previously commonly used spherical beads, this kind of encoded beads has a plane, so it is easier to aggregate and overlap, and it is more difficult to disperse.

Nicewarner-Pena et al. recently reported a method for synthesizing a code composed of submicron-scale stripes formed by various metals (Nicewarner-Pena et al., Science, 294(5540):137-41(2001)). Striped patterns can be formed on the base plate by sequentially depositing different metal ions on the base plate using electrochemical methods. Different stripes are composed of different metal materials and have different reflectivity. Such stripe codes can be identified by optical microscopes common in laboratories. In the biological analysis of DNA and protein, it is often necessary to detect the analytes that are affinity-bound to the beads by means of fluorescence, and the above-mentioned coding readout mechanism will not interfere with the detection of fluorescence.

A system that combines the advantages of both planar arrays and encoded microparticles could overcome the limitations of existing approaches. Illumina has already made such an attempt. They proposed a method of using microbeads and etched glass fibers to make arrays (Ferguson et al. High-density fiber-optic DNA random microspherearray. Anal. Chem., 72:5618-5624 (2000)). However, Illumina's approach using fluorescently encoded oligonucleotide-based beads is still limited by the number of different beads that can be obtained. BioArraySolutiohs uses the LEAPS (Light-controlled Electrokinetic Assembly ofParticles near Surfaces) method to form a bead array on the surface (WO 97/40385). However, this LEAPS approach is still prone to the problem that plagues most bead-based techniques, namely the limited number of types of codes that can be obtained.

Therefore, there is an urgent need to develop a micro-device that combines the advantages of micro-processed particles and planar discrete arrays to meet the needs of production and practice.

Contents of the invention

The object of the present invention is to provide a micro device.

The micro-device provided by the present invention includes: a) a magnetizable base material; b) an optically identifiable coding pattern fabricated on the base material; c) a magnetization axis.

In a certain practical application, the coding portion of the microdevice of the present invention is not realized by platinum, palladium, nickel, cobalt, silver, copper or gold; in another practical application, the composition material of the microdevice of the present invention does not include Platinum, palladium, nickel, cobalt, silver, copper or gold.

The microdevices can be fabricated using any suitable magnetizable material. For example, a paramagnetic substance, a ferromagnetic substance, or a ferrimagnetic substance may be used. The micro-device can also be made of suitable metal materials, for example, metal elements of the transition group can be used: iron, nickel, copper, cobalt, tungsten, tantalum, zirconium or alloy substances thereof, such as cobalt-tantalum-zirconium alloy ( CoTaZr alloy). In practical applications, metal oxides are used as magnetic substances.

The micro-device also includes a substrate made of non-magnetizable material. These non-magnetizable materials can be any suitable materials such as silicon materials (silicon, silicon dioxide, silicon nitride), plastics, glass, ceramics, rubber, polymers, alumina, aluminum, gold, titanium, etc. or their Complex. The magnetizable substrate material may be attached to the substrate in any suitable manner. For example, the magnetizable base material may be a part of the base, or may be connected to the base by deposition, bonding and the like.

The substrate may be a multi-layer structure, such as 3 layers, 4 layers or more layers. For example, the substrate can be a three-layer structure, the top layer and the bottom layer can be made of the same material, such as silicon dioxide (or glass), and the middle layer is made of magnetic material. Of course, the top and bottom layers can also be made of different materials.

The surface of the substrate can be either hydrophobic or hydrophilic. The substrate may be formed in any suitable shape, such as sphere, square, rectangle, triangle, disc, cube, parallelepiped, cone, cylinder, prism or other regular or irregular shapes. The substrate can take any suitable size, for example the thickness of the substrate can be between 0.1 micron and 500 microns; preferably between 1 micron and 200 microns; most preferably between 1 micron and 50 microns between. For example, the substrate can be rectangular with an area between 10 square microns and 1,000,000 square microns (1000 microns by 1000 microns). The substrate may also be circular with a diameter between 10 microns and 500 microns. The substrate can also be a cubic structure with side lengths ranging from 10 microns to 100 microns. The substrate can also be of an irregular shape with feature sizes between 1 micron and 500 microns. The substrate may include silicon layers, metal layers, and polymer layers. The substrate may also include a silicon layer and a metal layer, such as an aluminum layer. Preferably, the metal layer includes magnetic substances such as nickel or CoTaZr (Cobalt-Tantalum-Zirconium) alloy.

Any light-recognizable property can be used as a feature of the coded pattern. For example, the optically recognizable pattern can be given by the composition of the substance itself: holes punched on the base material; other materials with different optical properties from the base are fixed on the base material in a certain way. The coding method of the optically identifiable coding pattern can be based on the substrate itself or the hole punched on the substrate or the pattern, quantity, position distribution, optical properties, material composition or substance fixed or placed on the substrate material. A combination of the above methods. For example, the substrate can be a 4-layer structure, the top layer and the bottom layer can be made of the same material, such as silicon dioxide (or glass), and the middle layer is made of magnetic material, such as magnetic alloy. The other middle layer has an optically recognizable code as the code layer. Preferably, the magnetic layer and the code layer do not overlap each other, at least not completely, so as not to interfere with the optical detection of the code on the code layer. Of course, the top layer and the bottom layer can also be made of different materials. The code can be numbers, letters, structural patterns, one-dimensional or two-dimensional barcodes and the like.

The microdevices may have only one optically recognizable coding pattern, but the microdevices may also have multiple photorecognizable coding patterns, for example, a set of holes punched in the substrate or a set of A code composed of substances having different optical properties from the substrate arranged on the substrate.

In order to facilitate the optical analysis of the optical coding pattern, the micro-device preferably has a mark for positioning. For example, for disc-shaped micro-devices, when the micro-devices are placed flat, it is difficult to distinguish the front or back of the micro-devices, which will cause difficulties in identification. And just can determine which side is the front side that has coding pattern by positioning mark.

The photorecognizable coded pattern may be fabricated on the substrate using any suitable technique. For example, micromachining techniques can be used. Suitable microfabrication techniques include mask etching techniques such as photolithography, electron beam etching and X-ray etching (see World Patent WO 96/39937 and U.S. Patent Nos. 5,651,900, 5,893,974 and 5,660,680). Micromachining technology can be used to directly produce required coding patterns on the substrate, such as numbers, letters, structural patterns, one-dimensional or two-dimensional barcodes. Figure 8 gives examples of coding for some such microdevices. The picture shows the microdisk that has been fabricated but not yet removed from the silicon wafer. The magnification is about 400 times. A is a microdisc with rectangular ferrite bars and a 2D matrix code; B is a microdisc with rectangular but sharp-ended ferrite rods and a 3-character code; C is a rectangular ferrite bar but with The micro-disc at the end of the 3 ends, the encoding method is a 1D barcode; D is a micro-disc with a rectangular magnetic rod and a 4-character code.

If a material having different optical properties than the substrate material is used to make the coded pattern, this material may be arranged/fixed on the substrate using any suitable method. For example, sputtering or vapor deposition techniques can be used. This material can be arranged or immobilized on the substrate directly or via some linker, eg a cleavable linker. The required coding patterns, such as numbers, letters, structural patterns, one-dimensional or two-dimensional barcodes, can be directly produced on the substrate by using micro-processing technology. Such materials can be arranged or immobilized on the substrate by covalent or non-covalent linkages. This material can be arranged or immobilized on the substrate by specific or non-specific binding.

Any suitable optical marker can be used in the present invention. For example, such optical markers can be optically recognizable coding patterns composed of metal films made of metals such as copper, aluminum, gold and platinum, and these coding patterns can be numbers, letters, structural patterns, one-dimensional or two-dimensional dimension barcode. Such optical markers may be fluorescent substances, light-scattering detectable microparticles (see US Patent No. 6,214,560), quantum dots (US Patent No. 6,252,664), etc. as the above-mentioned optical markers.

Suitable quantum dot materials can be used with the present invention. For example, the quantum dots may be a cadmium-X (Cd-X) core structure, where X represents selenium (Se), sulfur (S) or tellurium (Te). The quantum dots can be further wrapped with a layer of inorganic substances for passivation. The wrapping layer has a Y-Z structure, wherein Y represents cadmium or zinc, and Z represents sulfur or selenium. Preferably, the structure of the quantum dot is a cadmium-X (Cd-X) core structure, wherein X represents selenium (Se), sulfur (S) or tellurium (Te); and a Y-Z type structure, wherein Y represents cadmium or It is zinc, and Z represents sulfur or selenium; in addition, the outer layer is wrapped with a layer of trialkylphosphine oxide to obtain quantum dots with a CdX core/YZ shell structure.

The quantum dots with the CdX core/YZ shell structure can adopt appropriate methods to make the produced quantum dots water-soluble. One approach is to replace the outer wrapping of the quantum dots of the CdX core/YZ shell structure with a layer of water soluble material. For example, mercaptocarboxylic acids can be used in place of trialkylphosphine oxides in the outer layer of quantum dots. The specific operation is to use a large amount of mercaptocarboxylic acid to treat the quantum dots. _Quantum dots can also be treated with a CHCl solution containing a large amount of mercaptocarboxylic acids (see Chan and Nie, Science, 281:2016-2018 (1998)). The mercapto groups in the coating molecules can form a relatively stable Cd(Zn)-S bond with the quantum dots in solution. Another treatment method to make the quantum dots with the CdX core/YZ shell structure water-soluble is to silanize the quantum dots (see Bruchez et al., Science, 281: 2013-2015 (1998)). Silanizing the surface of nanomaterials can make the material water-soluble, and the silanized surface can be further chemically modified. In general, these “water-soluble” quantum dot materials still need to be further functionalized to make the quantum dots stable enough in aqueous solution and exposure to light and air (oxygen) for the application in fluorescence detection systems ( See US Patent No. 6,114,038). Because of the unique fluorescent properties of quantum dot materials (such as, but not limited to, high quantum yield, not easy to be photobleached, stable in complex aqueous solution systems, etc.), water-soluble quantum dot materials are very sensitive in detection . Another advantage of quantum dot materials is that a series of quantum dot materials can be produced, and they can emit different fluorescences under the same excitation light.

Quantum dots suitable for use in the present invention can be of any suitable size. For example, quantum dots can range in size from 1 nanometer to 100 nanometers.

The micro-device used in the present invention may contain only one kind of quantum dots, or may contain multiple kinds of quantum dots. For example at least two quantum dots with different sizes and different fluorescence can be included.

The micro-devices used in the present invention may contain only one kind of optical markers, or may contain multiple kinds of optical markers. For example at least two different optical markers may be included.

The microdevices of the present invention may contain electrically conductive or dielectrically polarizable substances. These electrically conductive or dielectrically polarizable substances added to microdevices can change the electrical or dielectric properties of the entire microdevices, thereby affecting the interaction between the external electric field and the microdevices, and affecting the external electric field received by the microdevices Inductive forces (such as dielectrophoretic forces, traveling wave dielectrophoretic forces, etc.).

In order to achieve the purpose of using the micro-device for separation, manipulation or detection, it is necessary to properly select parameters such as material, composition, structure, and size of the micro-device. For example, the microdevices can be used to separate specific molecules from mixtures. If dielectrophoretic forces are used for separation, the microdevices should have certain dielectric properties. If magnetic force is used for separation or manipulation, the fabrication materials of the micro-devices should contain magnetic substances, such as ferromagnetic or ferrimagnetic materials.

The micro-device may carry binding substances for binding entity molecules to be separated/manipulated/detected. Preferably, the conjugate itself can specifically recognize and bind to the corresponding entity molecule. In the description of the present invention, where a combination is mentioned, a combination stands for a combination of a combination of a combination and a microdevice. For example, if the description mentions the complex formed by the conjugate and the entity molecule, it actually refers to the complex formed by the conjugate and the entity molecule connected to the micro-device.

Here, the conjugate can use any suitable substance (see pending U.S. Patent Application Serial Nos. 09/636,104, filed August 10, 2000 and 09/679,024, filed October 4, 2000) ). Conjugates can be cells such as animal, plant, fungal or bacterial cells; organelles such as nucleus, mitochondria, chloroplasts, ribosomes, endoplasmic reticulum, Golgi apparatus, lysosomes, proteasomes, vesicles, vacuoles or microbodies; viruses, Microparticles or their polymers or composites. Conjugates can also be molecules immobilized on the surface of a microdevice. For example, antibody molecules bound to the surface of a micro-device. Microdevices bound to antibody molecules can be used to capture target proteins from mixtures of molecules, and target cells from mixtures of cells. Oligo-dT (for example, a nucleic acid molecule consisting of 25 Ts) can also be immobilized on the surface of the microdevice. Microdevices incorporating Oligo-dT can be used to isolate mRNA from molecular mixtures. For capturing DNA molecules, other conjugates can also be used. Nucleic acid fragments, such as DNA, RNA; peptide nucleic acid sequences can be used to hybridize with target nucleic acid (DNA, RNA) or peptide nucleic acid. The surface of the micro-device can be connected or combined with molecules or functional groups, etc., so that the surface of the micro-device forms a functional layer that can perform various chemical/biochemical/biological reactions . Through various reactions, the entity molecules can be combined on the surface of the micro-device, so that the manipulation, separation or detection of the entity molecules can be carried out in the next step through the operation of the micro-device. The functional layer on the surface of the micro-device also makes it possible to carry out the synthesis reaction directly on the surface of the micro-device. Such synthesis may be of synthetic nucleic acids (such as DNA and RNA), but also of synthetic polypeptides or proteins. The surface of such a functional layer includes, but is not limited to, modified carboxyl, amino, hydroxyl, mercapto, epoxy, ester, alkenyl, alkynyl, alkyl, aromatic, aldehyde, ketone and other groups or are the surfaces of their derivatives.

The choice of the microdevices is related to the specific means of separation, manipulation and detection. For example, when dielectrophoretic forces are used to separate target molecules from a mixture, the dielectric properties of the microdevice or the conjugate must be substantially different from the other molecules in the mixture so that when the conjugate binds to the target entity , the entity molecule-conjugate-microdevice complex can be selectively manipulated by dielectrophoresis. For example, when it is necessary to separate cancer cells from a mixture, the dielectric properties of cancer cells are very close to normal cells, and almost all cells have similar dielectric properties, such as negative dielectrophoresis. In this case, the microdevices or conjugates should be made of materials that are more easily polarized than the medium to exhibit forward dielectrophoresis. In this way, the microdevice-conjugate-cancer cell complex can be selectively manipulated using positive DEP forces, while other cells are subjected to negative DEP forces.

Microdevices can contain only one conjugate or multiple conjugates for high-throughput separation, manipulation and detection.

Because the micro-device contains magnetizable substances, the micro-device, the micro-device-solid molecule complex or the micro-device-conjugate-solid molecule complex can rotate, move or be manipulated under the action of a magnetic field force. The magnetic field force refers to the force that particles and other solid molecules are subjected to due to the existence of a magnetic field. In general, for sufficient magnetic force to manipulate a particle, the particle must be magnetic (for example, paramagnetic or ferromagnetic), or capable of being magnetized or polarized. Let us consider an example of a typical magnetic particle composed of superparamagnetic matter, when this particle is placed in the magnetic field B, the particle will produce a magnetic dipole μ,

$\stackrel{<span\; class="notranslate">\; \&OverBar;</span>}{\mathrm{<span\; class="notranslate">\; \&mu;</span>}}<span\; class="notranslate">\; =</span>{\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}<span\; class="notranslate">\; (</span>{\mathrm{<span\; class="notranslate">\; \&chi;</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}<span\; class="notranslate">\; -</span>{\mathrm{<span\; class="notranslate">\; \&chi;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; )</span>\frac{\stackrel{<span\; class="notranslate">\; \&OverBar;</span>}{\mathrm{<span\; class="notranslate">\; B</span>}}}{{\mathrm{<span\; class="notranslate">\; \&mu;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}}<span\; class="notranslate">\; ,</span>$

$<span\; class="notranslate">\; =</span>{\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}<span\; class="notranslate">\; (</span>{\mathrm{<span\; class="notranslate">\; \&chi;</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}<span\; class="notranslate">\; -</span>{\mathrm{<span\; class="notranslate">\; \&chi;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}<span\; class="notranslate">\; )</span>{\stackrel{<span\; class="notranslate">\; \&OverBar;</span>}{\mathrm{<span\; class="notranslate">\; h</span>}}}_{\mathrm{<span\; class="notranslate">\; m</span>}}$

Among them, V _p is the volume of the particle, χ _p and χ _m are the magnetic susceptibility coefficients of the particle and its surrounding medium, μ _m is the magnetic permeability of the medium, and H _m is the magnetic field strength. The magnetic field force F _magnetic on the particle can be obtained by the instantaneous magnetic dipole and magnetic field gradient ${\stackrel{\&OverBar;}{f}}_{\mathrm{magnetic}}=-0.5V_p({\mathrm{\&chi;}}_p-{\mathrm{\&chi;}}_m)\stackrel{\mathrm{\&omega;}}{h_m}\&CenterDot;\&dtri;{\stackrel{\mathrm{\&rho;}}{B}}_m,$

”分别指的是点乘和梯度运算。很明显是否有磁场力加在微粒上取决于粒子和其周围介质间的磁化系数。一般粒子是被悬于非磁性的液体介质中(其磁化系数接近于零)，因此必须利用磁性微粒(其磁化系数远大于0)。在磁场力与粘附力达到平衡时粒子的速度v_particle可由下式给出Where the symbols "·" and "

"Refer to the dot product and gradient operation respectively. Obviously whether there is a magnetic field force on the particle depends on the magnetic susceptibility between the particle and its surrounding medium. Generally, the particle is suspended in a non-magnetic liquid medium (the magnetic susceptibility is close to zero), so magnetic particles (its magnetic susceptibility coefficient is much greater than 0) must be used. When the magnetic field force and the adhesion force reach a balance, the particle velocity v _particle can be given by the following formula

${\mathrm{<span\; class="notranslate">\; v</span>}}_{\mathrm{<span\; class="notranslate">\; particles</span>}}<span\; class="notranslate">\; =</span>\frac{{\stackrel{<span\; class="notranslate">\; \&OverBar;</span>}{\mathrm{<span\; class="notranslate">\; f</span>}}}_{\mathrm{<span\; class="notranslate">\; magnetic</span>}}}{{\mathrm{<span\; class="notranslate">\; 6</span>}\mathrm{<span\; class="notranslate">\; \&pi;r\&eta;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}}$

where r is the radius of the particle and _ηm is the viscosity of the surrounding medium. Therefore, in order to obtain a sufficiently large magnetic manipulation force, the following factors must be considered: (1) the magnetizability of the particles should be optimized, (2) the magnetic field strength should be maximized, and (3) the field strength gradient of the magnetic field should be optimized.

Paramagnetic particles refer to those particles that can induce magnetic dipoles under the action of an external magnetic field, and when the magnetic field is removed, the magnetic dipoles can return to zero. In the application of the present invention, commercially available paramagnetic particles or magnetic particles can be used. The size of these particles generally ranges from submicron (50 nm to 0.5 nm) to tens of microns. These particles have different structures and compositions. One type of particle is made of a ferromagnetic material surrounded by a polymer layer, such as polystyrene. Another type of microparticles is filled with ferromagnetic nanoparticles in the pores of porous beads (such as polystyrene beads). The surfaces of these two types of microparticles can be unmodified polystyrene or modified to facilitate the attachment of various molecules. There is also a kind of particle, which is made by uniformly doping ferrimagnetic substances in the polymerization step of the production process. Thus, the microdevices of the present invention contain these paramagnetic or magnetic particles, so that the microdevices contain components that can be magnetized.

The microdevices can be fabricated using any suitable magnetizable material. For example, paramagnetic substances, ferromagnetic substances, ferrimagnetic substances or superparamagnetic substances can be used to directly connect or fabricate on micro-devices. Micro-devices can also be made of suitable metal materials, for example, metal elements of the transition group can be used: iron, nickel, copper, cobalt, tungsten, tantalum, zirconium or alloys thereof, such as cobalt-tantalum-zirconium alloy (CoTaZr alloy ). However, various methods are used, such as electroplating (for example to make iron-nickel alloy) or sputtering (for example to make cobalt-tantalum-zirconium alloy) to make the magnetizable substance. In the U.S. patent application "Individually Addressable Micro-ElectromagneticUnit Array Chips in Horizontal Configurations" (Wu et al., application number 09/685,410, application date October 10, 2000), a variety of methods for making magnetizable substances (paramagnetic substances) are given. and ferromagnetic substances, etc.).

The rotation or manipulation of magnetic particles, micro-device, micro-device-solid molecule complex or micro-device-conjugate-solid molecule complex needs to generate a magnetic field first. Particles that can be manipulated by magnetic force should have a large magnetic susceptibility. In addition, the particles should have no or a small amount of remanence after the magnetic field is removed. Micro-electromagnetic units can be used to generate the required magnetic fields described above. When a current is applied to the micro-electromagnetic unit, the electromagnetic unit can induce a magnetic field. By controlling the current applied to the micro-electromagnetic unit, the distribution of the magnetic field can be controlled. The structure and size of the micro-electromagnetic unit can be designed according to the required magnetic field. The manipulation of magnetic particles includes linear motion, focusing motion, induced motion, etc. The movement of magnetic particles in a magnetic field is called "magnetophoresis". Theory and examples of magnetophoresis for cell separation and other applications can be found in many literatures. Such as: Magnetic Microspheres in Cell Separation, by Kronick, P.L. in Methods of Cell Separation, Volume 3, edited by N.Catsimpoolas, 1980, pages 115-139; Use of magnetic techniques for the isolation of cells, by Safarik I.And , inafarikovaM J.of Chromatography, 1999, Volume 722(B), pages 33-53; A fully integrated micromachined magnetic particle separator, by Ahn C.H.et al., in J.of Microelectromechanical systems, 1996, Volume 5, pages 151-157.

The microdevice may also comprise a unit that facilitates or enables manipulation of the microdevice/solid molecule-microdevice complex. The unit can be made of conductive material to be manipulated by dielectrophoretic forces; material with high/low acoustic damping properties to be manipulated by acoustic field forces; and charged material to be manipulated by electrostatic forces. This unit can be a cell, organelle, virus, particle, molecular aggregate or their complex. In addition, combinations mentioned in pending U.S. Patent Application Serial No. 09/636,104 (filed August 10, 2000) can also be used as the above-mentioned units. Such substances can be, but are not limited to, substances with special physical and chemical properties produced by deposition or other processes. Such substances can be metal films made of metals such as gold, chromium, titanium or platinum, which can not only be used as constituent materials of micro-devices, but also can increase the conductivity of micro-devices. Such substances can also be insulating substances such as plastic polymers such as polystyrene, which can not only be used as constituent materials of micro-devices, but also can reduce the electrical conductivity of micro-devices.

The units in the above-mentioned micro-device can facilitate the manipulation of the micro-device/solid molecule-micro-device complex through a suitable physical field, or enable the micro-device/solid molecule-micro-device complex to be manipulated using a suitable physical field become possible. Such physics can be found in pending U.S. Patent Application Serial No. 09/636,104 (filed August 10, 2000). For example, it can be dielectrophoretic force, traveling wave dielectrophoretic force, magnetic field force (in the magnetic field generated by ferromagnetic material or micro-electromagnetic unit), acoustic field force (in standing wave field or traveling wave field), static force Electric force (such as in a DC electric field), mechanical force (such as liquid shear force), optical radiation force (generated by optical pressure of optical tweezers), etc.

Dielectrophoresis is the movement of polarized particles (micro-device, micro-device-solid molecule complex or micro-device-conjugate-solid molecule complex) in a non-uniform amplitude electric field. When a particle is placed in an electric field, if there is a certain difference in the dielectric properties of the particle and its surrounding medium, the particle will be polarized. In this way, charges are induced at the particle-medium interface. If the applied electric field is a non-uniform AC electric field, then the induced charge of the particles interacts with the applied electric field, so that the net force on the particles is not zero, so that the particles move to the region of strong or weak field. The net force on the particle is called DEP force, and the motion of the particle is called DEP. The dielectrophoretic force on a particle is determined by the dielectric properties of the particle itself, the dielectric properties of the medium around the particle, and the frequency and amplitude distribution of the applied electric field.

Traveling wave dielectrophoresis is similar to the above-mentioned dielectrophoresis. The traveling wave electric field acts on the polarized charges on the particles caused by the electric field, so that the particles (microdevices, microdevices-solid molecular complexes or microdevices- Conjugates-entity molecule complexes) are subjected to corresponding forces, making the particles move along or against the transmission direction of the traveling wave electric field. The traveling wave dielectrophoretic force on a particle is determined by the dielectric properties of the particle itself, the dielectric properties of the medium around the particle, and the frequency, amplitude and phase distribution of the traveling wave electric field. The theory of dielectrophoresis and traveling wave dielectrophoresis and methods for manipulating particles using dielectrophoresis can be found in the following reference: "Non-uniform SpatialDistributions of Both the Magnitude and Phase of AC Electric Fields determine Dielectrophoretic Forces" Wang et al. Biochim. Biophys. Acta 1243: 185-194 (1995); "Dielectrophoretic Manipulation of Particles" Wang et al. IEEE Transaction on Industry Applications 33: 660-669 (1997); "Huang et al.J.Phys.D: Appl.Phys.26:1528-1535; "Positioning and manipulation of cells and microparticles using miniaturized electric fieldtraps and traveling waves" Fuhr et al.Sensors and Materials 7:131-146;" Dielectrophoretic manipulation of cells using spiral electrodes" Wang et al. Biophys. J.72: 1887-1899 (1997); "Separation of human breast cancer cells from blood by differential dielectric affinity" Becker et al. Proc. Natl. Acad. Sci. : 860-864 (1995). Manipulation of particles using dielectrophoresis and traveling wave dielectrophoresis includes concentration, aggregation, trapping, repulsion, suspension, separation, or linear or other directional motion of particles. Particles can be trapped and concentrated in specific areas of the reaction cell. Particles can be subdivided into subclasses at a fine level, and can also be transported over a certain distance. The electric field distribution required for the manipulation of specific particles can be designed through the relevant dielectrophoresis theory and electric field simulation methods. The distribution of the electric field in specific applications is determined by the size and geometry of the electrodes.

In a non-uniform electric field, the dielectrophoretic force F _DEPz on a particle with a radius r can be expressed as:

${\mathrm{<span\; class="notranslate">\; f</span>}}_{\mathrm{<span\; class="notranslate">\; DEPz</span>}}<span\; class="notranslate">\; =</span>{\mathrm{<span\; class="notranslate">\; 2</span>}\mathrm{<span\; class="notranslate">\; \&pi;\&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}{\mathrm{<span\; class="notranslate">\; <figure-callout\; id="3"\; label="r"\; filenames="C0312082100541.png"\; state="\{\{state\}\}">r</figure-callout></span>}}^{\mathrm{<span\; class="notranslate">\; 3</span>}}{\mathrm{<span\; class="notranslate">\; \&chi;</span>}}_{\mathrm{<span\; class="notranslate">\; DEP</span>}}<span\; class="notranslate">\; \&dtri;</span>{\mathrm{<span\; class="notranslate">\; <figure-callout\; id="2"\; label="E.\; rms"\; filenames="C0312082100541.png"\; state="\{\{state\}\}">E.</figure-callout></span><figure-callout\; id="2"\; label="E.\; rms"\; filenames="C0312082100541.png"\; state="\{\{state\}\}">\; </figure-callout>}}_{\mathrm{<span\; class="notranslate">\; </span>}}^{\mathrm{<span\; class="notranslate">rms</span>}}<span\; class="notranslate">\; \&Center\; Dot;</span>{\stackrel{\mathrm{<span\; class="notranslate">\; \&rho;</span>}}{\mathrm{<span\; class="notranslate">\; a</span>}}}_{\mathrm{<span\; class="notranslate">\; z</span>}}$

where E _rms is the root mean square value of the field strength, symbol is the symbol representing the gradient operation, and ε _m is the permittivity of the medium. _χDEP is the polarization factor of the particle, which can be expressed as:

${\mathrm{<span\; class="notranslate">\; \&chi;</span>}}_{\mathrm{<span\; class="notranslate">\; DEP</span>}}<span\; class="notranslate">\; =</span>\mathrm{<span\; class="notranslate">\; Re</span>}<span\; class="notranslate">\; (</span>\frac{{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}^{<span\; class="notranslate">\; *</span>}<span\; class="notranslate">\; -</span>{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}^{<span\; class="notranslate">\; *</span>}}{{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}^{<span\; class="notranslate">\; *</span>}<span\; class="notranslate">\; +</span>{\mathrm{<span\; class="notranslate">\; 2</span>}\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}^{<span\; class="notranslate">\; *</span>}}<span\; class="notranslate">\; )</span>$

Here Re refers to the real part of a complex number, the symbol ${\mathrm{\&epsiv;}}_x^{*}={\mathrm{\&epsiv;}}_x-j{\mathrm{\&sigma;}}_x/\left(2\mathrm{\&pi;f}\right)$ is the complex permittivity (where x=p corresponds to the particle and x=m corresponds to the medium). The parameters ε _p and σ _p are the effective permittivity and conductivity of the particle respectively (may be related to the external frequency). For example, a typical biological cell will have a frequency-dependent electrical conductivity and permittivity (induced at least in part by polarization of the cell's plasma membrane).

The above formula describing the dielectrophoretic force can also be expressed as

${\mathrm{<span\; class="notranslate">\; f</span>}}_{\mathrm{<span\; class="notranslate">\; DEPz</span>}}<span\; class="notranslate">\; =</span>{\mathrm{<span\; class="notranslate">\; 2</span>}\mathrm{<span\; class="notranslate">\; \&pi;\&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}{\mathrm{<span\; class="notranslate">\; <figure-callout\; id="3"\; label="r"\; filenames="C0312082100541.png"\; state="\{\{state\}\}">r</figure-callout></span>}}^{\mathrm{<span\; class="notranslate">\; 3</span>}}{\mathrm{<span\; class="notranslate">\; \&chi;</span>}}_{\mathrm{<span\; class="notranslate">\; DEP</span>}}{\mathrm{<span\; class="notranslate">\; V</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}\mathrm{<span\; class="notranslate">\; p</span>}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; z</span>}<span\; class="notranslate">\; )</span>{\stackrel{\mathrm{<span\; class="notranslate">\; \&rho;</span>}}{\mathrm{<span\; class="notranslate">\; a</span>}}}_{\mathrm{<span\; class="notranslate">\; z</span>}}$

Among them, p(z) is the distribution of the square of the electric field value of unit voltage excitation (voltage V=1V) on the electrode, and V is the applied voltage.

There are generally two forms of dielectrophoresis, positive dielectrophoresis and negative dielectrophoresis. In forward dielectrophoresis, the particles move to the strong field area under the action of dielectrophoretic force. In negative dielectrophoresis, the particles move to the weak field area under the action of dielectrophoretic force. The positive or negative dielectrophoresis of the particles is determined by the relative polarization of the particles and the medium.

Traveling wave dielectrophoretic force refers to the force experienced by solid molecules (particles or molecules) in a traveling wave electric field. The characteristic of traveling wave electric field is that the phase value of its AC electric field component is unevenly distributed.

Here, we analyze the traveling wave dielectric electric field force for an ideal traveling wave electric field. in traveling wave electric field ${\mathrm{E.}}_{\mathrm{TWD}}=\mathrm{E.}\cos \left(2\mathrm{\&pi;}(\mathrm{ft}-z/{\mathrm{\&lambda;}}_0)\right){\stackrel{\mathrm{\&rho;}}{a}}_x$ (For example, when an electric field in the x direction runs along the z direction), the dielectrophoretic force F _TWD dielectrophoretic force acting on a particle with a radius of r can be given by the formula $f_{\mathrm{TWD}}=-2{\mathrm{\&pi;\&epsiv;}}_m{\mathrm{<figure-callout\; id="3"\; label="r"\; filenames="C0312082100541.png"\; state="\{\{state\}\}">r</figure-callout>}}^3{\mathrm{\&zeta;}}_{\mathrm{TWD}}{\mathrm{E.}}^2\&Center\; Dot;{\stackrel{\mathrm{\&rho;}}{a}}_z$ where E is the field strength, ε _m is the dielectric constant of the medium, ζ _TWD is the polarization factor of the particle, and ζ _TWD is given by ${\mathrm{\&zeta;}}_{\mathrm{TWD}}=\mathrm{Im}\left(\frac{{\mathrm{\&epsiv;}}_p^{*}-{\mathrm{\&epsiv;}}_m^{*}}{{\mathrm{\&epsiv;}}_p^{*}+{2\mathrm{\&epsiv;}}_m^{*}}\right),$ where Im refers to the imaginary part of the complex number, the symbol ${\mathrm{\&epsiv;}}_x^{*}={\mathrm{\&epsiv;}}_x-{\mathrm{j\&sigma;}}_x/2\mathrm{\&pi;f}$ is the composite permittivity (of the particle x=p and the medium x=m), the parameters ε _p and σ _p are the effective permittivity and the conductivity of the particle, respectively, these parameters are frequency dependent.

Particles with different dielectric properties (as defined by dielectric constant and conductivity), such as biological cells, will exhibit different dielectrophoretic forces. For TWDEP manipulation of particles (including biological cells), the TWDEP force loaded on a particle with a diameter of 10 microns can vary between 0.01-10000 pN.

A traveling wave electric field can be generated by applying a suitable AC signal to the microelectrodes arranged on the chip. In order to generate a traveling wave electric field, at least three sets of electrical signals with different phase values should be applied to the electrodes. In an example of a traveling wave electric field, four electrical signals with different phase values (0, 90, 180, and 270 degrees respectively) are used and applied to four linear parallel electrodes arranged on the chip surface, such that Four electrodes can be used as a basic repeating unit for chip design. According to the needs of the application, more than two such units can be arranged to generate a traveling wave electric field above or near the electrodes. As long as the electrode units are arranged in a certain order in space, the external signals of different phases can generate a traveling wave electric field in the vicinity of the electrodes.

The dielectrophoretic force and traveling wave dielectrophoretic force on particles (microdevice, microdevice-solid molecule complex or microdevice-conjugate-solid molecule complex) are not only related to the electric field distribution (electric field amplitude , phase, frequency, etc.), and is related to the dielectric properties of particles and media. If the particle is more polarized than the medium (for example, at a certain frequency, the particle has greater conductivity and polarizability), the particle will move to the strong field area under the action of the forward dielectrophoretic force. If the particle is less polarized than the medium, the particle moves to the weak field region under the action of negative dielectrophoretic force. The positive or negative dielectrophoresis of the particles is determined by the relative polarization of the particles and the medium. In traveling wave dielectrophoresis, the particles are subjected to dielectrophoretic force, and according to the difference of the polarization factor ζ _TWD , the particles can move along or against the direction of the traveling wave electric field. The following documents give some basic theories about dielectrophoresis and traveling wave dielectrophoresis: Huang, et al., J. Phys.D: Appl. Phys. 26: 1528-1535 (1993); Wang, et al. , Biochim. Biophys. Acta. 1243: 185-194 (1995); Wang, et al., IEEE Trans. Ind. Appl. 33: 660-669 (1997).

Particles (microdevices, microdevice-entity molecule complexes or microdevice-associated-entity molecule complexes) can be manipulated by acoustic field radiation forces, for example in an acoustic field. If a standing wave sound field is generated in space by a sound field source and its reflected waves, the particles in it can be affected by the radiation force of the sound field according to the difference between the sound damping coefficient of itself and the surrounding medium. Acoustic damping is related to the density of the material itself, and is the speed at which sound waves travel through the material. If the particle is more acoustically damped than the surrounding medium, it experiences acoustic field radiation forces directed at the nodes of the standing wave acoustic field. When the sound field distribution is different, the particles are subjected to different sound field radiation forces.

A piezoelectric material can be used to generate an acoustic field. When an electrical signal of a suitable frequency is applied to the piezoelectric material, the piezoelectric material can generate mechanical vibrations and transmit them to the medium around the material. Piezoceramic is one such piezoelectric material. Microelectrodes can be fabricated on the piezoceramic to apply an electrical signal to cause the piezoceramic to generate an acoustic field. According to different application needs, microelectrodes of different sizes and geometries can be designed. In order to generate a standing wave sound field, reflective walls are also required. Sound fields of different frequencies can be used in the present invention, for example from a few kHz to several hundred MHz. In addition to standing wave sound fields, non-standing wave sound fields, such as traveling wave sound fields, can also be used. The traveling wave sound field can also make the particles in it subject to force, see "Acoustic radiation pressure on a compressible sphere, by K. Yoshioka and Y. Kawashima in Acustica, 1955, Vol.5, pages 167-173". The particles are not only affected by the sound field, but also by the force of the surrounding fluid medium, and the fluid medium moves under the action of the traveling wave sound field. Using acoustic fields, microparticles can be enriched, focused, trapped, lifted, and transported in a microfluidic environment. Another mechanism by which forces are exerted on particles in an acoustic field is through acoustic field-induced convection of the fluid medium. Such convection is determined by the distribution of the sound field itself, the nature of the fluid, and the volume and structure of the cavity in which the fluid resides. Such convection of the fluid can exert a force on the particles in it so as to realize the manipulation of the particles. This manipulation can be used to facilitate mixing of the fluid and the particles therein. In the present invention, such convection can be used to promote the mixing of the conjugates attached to the micro-device and the entity molecules in the fluid, and enhance the interaction between the conjugates and the entity molecules.

An ultrasonic standing wave can be generated by applying an AC signal to a piezoelectric transducer. For example, assume that a standing wave is established in a liquid in a specific direction (eg, the Z direction). The change of the standing wave in the Z direction can be expressed as follows:

Δp(z)=p ₀ sin(kz)cos(ωt)

Where Δp represents the sound field pressure in the Z direction, p ₀ represents the sound field pressure intensity, k represents the wave number (2π/λ, where λ is the wavelength), z represents the distance to the pressure node, ω represents the angular frequency, and t represents time. According to the theory proposed by Yoshioka and Kawashima in 1955, in a static standing wave field, the sound field radiation force acting on a spherical particle can be expressed by the following formula, see K.Yoshioka and Y.Kawashima in Acustica, 1955, Vol.5 , pages 167-173; "Acoustic radiation pressure on a compressible sphere and electrostatic force by Yasuda K. et al. in Jpn. J. Appl. Physics, 1996, Volume 35, pages 3295-3299":

${\mathrm{<span\; class="notranslate">\; f</span>}}_{\mathrm{<span\; class="notranslate">\; acoustic</span>}}<span\; class="notranslate">\; =</span><span\; class="notranslate">\; -</span>\frac{\mathrm{<span\; class="notranslate">\; 4</span>}\mathrm{<span\; class="notranslate">\; \&pi;</span>}}{\mathrm{<span\; class="notranslate">\; 3</span>}}{\mathrm{<span\; class="notranslate">\; r</span>}}^{\mathrm{<span\; class="notranslate">\; 3</span>}}\mathrm{<span\; class="notranslate">\; k</span>}{\mathrm{<span\; class="notranslate">\; E.</span>}}_{\mathrm{<span\; class="notranslate">\; acoustic</span>}}\mathrm{<span\; class="notranslate">\; A</span>}\mathrm{<span\; class="notranslate">\; sin</span>}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; 2</span>}\mathrm{<span\; class="notranslate">\; kz</span>}<span\; class="notranslate">\; )</span>$

Where r is the radius of the particle, E _acoustic is the average energy density of the sound field, A is a constant, expressed by the following formula:

$\mathrm{<span\; class="notranslate">\; A</span>}<span\; class="notranslate">\; =</span>\frac{{\mathrm{<span\; class="notranslate">\; 5</span>}\mathrm{<span\; class="notranslate">\; \&rho;</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}<span\; class="notranslate">\; -</span>{\mathrm{<span\; class="notranslate">\; 2</span>}\mathrm{<span\; class="notranslate">\; \&rho;</span>}}_{\mathrm{<span\; class="notranslate">\; <figure-callout\; id="2"\; label="m"\; filenames="C0312082100541.png"\; state="\{\{state\}\}">m</figure-callout></span>}}}{{\mathrm{<span\; class="notranslate">\; 2</span>}\mathrm{<span\; class="notranslate">\; \&rho;</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}<span\; class="notranslate">\; +</span>{\mathrm{<span\; class="notranslate">\; \&rho;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}}<span\; class="notranslate">\; -</span>\frac{{\mathrm{<span\; class="notranslate">\; \&gamma;</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}}{{\mathrm{<span\; class="notranslate">\; \&gamma;</span>}}_{\mathrm{<span\; class="notranslate">\; m</span>}}}$

Among them, ρ _m and ρ _p represent the density of particles and medium, and γ _m and γ _p represent the acoustic impedance of particles and medium. A here represents the polarization parameter of the sound field. The acoustic impedance is defined as the product of the density of the material (ρ _m and ρ _p represent the density of particles and media) and the sound velocity (C _m and C _p represent the sound speed in media and particles) (γ _m = ρ _m · C _m and γ _p = ρ _p · C _p ).

When A>0, the particle moves to the pressure node (z=0) of the standing wave.

When A<0, the particle moves away from the pressure node (z=0) of the standing wave.

Obviously, when particles with different densities and acoustic impedances are placed in the standing wave field, they are subject to different acoustic field radiation forces. For example: According to the established sound field energy distribution, the sound field radiation force experienced by a particle with a diameter of 10 microns varies from 0.01 to 1000pN.

Piezoelectric transducers are made of piezoelectric materials. When a mechanical force is applied to the piezoelectric material from the outside to deform it, the piezoelectric material generates an electric field (this is called the piezoelectric effect or the generator effect). Conversely, applying an electric field to a piezoelectric material produces a mechanical stress (this is called the electrostriction or motor effect). The piezoelectric effect was discovered in 1880 by Pierre Curie and his brother Jacques. The reason for the explanation is that the displacement of ions leads to the electrical polarization of the structural units of the material. When an electric field is applied, the ions are displaced by electrostatic forces, resulting in mechanical denaturation of the entire material. In this way, in a sound field-field flow separation or sound field-electrophoresis-field flow separation or sound field-dielectrophoresis-field flow separation device, an AC signal is applied to the piezoelectric transducer, and the vibration generated on the transducer is coupled to the reaction in the liquid in the cell, causing sound waves to be generated in the reaction cell. Such sound waves may have traveling and standing wave components.

Particles (microdevices, microdevice-solid molecule complexes, or microdevice-conjugation-solid molecule complexes) can be manipulated in a DC electric field. A DC electric field can exert an electrostatic force on charged particles. The electrostatic force is related to the quantity and polarity of the charges carried by the particle itself, and also related to the field strength and direction of the electric field itself. Particles with positive charge will move towards the electrode under the action of negative potential; particles with negative charge will move towards the electrode under the action of positive potential. When designing microelectrodes in microfluidic devices, consideration must be given to the correct spatial distribution of the electric field. In a DC electric field, particles in microfluidic devices can be enriched, focused, and transported. The surface of the electrode should also be covered with a suitable insulating layer to prevent or minimize unwanted surface electrochemical phenomena, thereby protecting the surface of the electrode.

由下式给出：The DC electric field force acting on a particle in the electric field F _E

is given by:

${\mathrm{<span\; class="notranslate">\; f</span>}}_{\mathrm{<span\; class="notranslate">\; E.</span>}}<span\; class="notranslate">\; =</span>{\mathrm{<span\; class="notranslate">\; Q</span>}}_{\mathrm{<span\; class="notranslate">\; p</span>}}{\mathrm{<span\; class="notranslate">\; E.</span>}}_{\mathrm{<span\; class="notranslate">\; z</span>}}{\stackrel{\mathrm{<span\; class="notranslate">\; \&rho;</span>}}{\mathrm{<span\; class="notranslate">\; a</span>}}}_{\mathrm{<span\; class="notranslate">\; z</span>}}$

where _Qp is the effective charge on the particle. The direction of the DC electric force acting on the charged particle is determined by the polarity of the charge on the particle and the direction of the applied electric field.

If a temperature gradient is established within the medium, such a temperature gradient will cause the flow of the medium, thereby forming a velocity field in the medium. The particles (micro-device, micro-device-solid molecule complex or micro-device-combination-solid molecule complex) are subjected to force. The flow part of this medium is caused by the fact that the medium tends to reach thermal equilibrium due to thermal diffusion. In addition, for aqueous solutions, the temperature distribution of the solution tends to be consistent with the corresponding density distribution. Such a density distribution can also lead to the medium flow to balance. Media flow caused by temperature gradients is sometimes referred to as thermal convection. Thermal convection can occur throughout a medium on a relatively large scale, and it can be used to facilitate mixing of liquids or as a force to bring molecules from distant locations to a specific reaction site.

Temperature gradient profiles can also be established in chip-based reactors or instruments by installing heating and/or cooling elements in the chip structure. Such a heating element can be a simple Joule heating resistor, which can be machined into the chip. Taking a heater with an impedance of 10 ohms as an example, passing a current of 0.2A will generate 0.4W Joule heat energy. If the coil is installed in an area smaller than 100 square microns, it will effectively increase the local temperature and in the A temperature gradient is created in the medium. Likewise, the cooling element may be a semiconductor Peltier element that causes the ambient temperature to drop when current is passed through it.

In practice, a chip may have an array of selectable heating elements that must be placed in a certain sequence or structured so that a temperature gradient is established when each, some, or all of the elements are activated This produces ideal heat convection and thus establishes a velocity field in the medium introduced into the device consisting of chips. For example, when a heating element is activated or energized, an increase in the temperature of the adjacent medium around this element will create a local temperature gradient, generating thermal convection. In another practical application, a chip may include multiple, interconnected heating cells so that the cells can be switched on and off synchronously. In yet another practical application, the chip may contain only a heating element, which raises the local temperature when current is applied, creating thermal convection in the medium. Likewise, the chip may contain an array of individually strobeable cooling elements, or a single cooling element.

In addition to the physical forces mentioned above, other physical forces can also be used in the present invention. For example, mechanical force (fluid force) can be used to transport particles (microdevice, microdevice-solid molecule complex or microdevice-associated-solid molecule complex). Optical field forces or optical radiation forces have been used in "optical tweezers" to aggregate, trap, levitate and manipulate particles. When a material with a refractive index different from its surrounding medium (such as particles, etc.) is placed in a gradient light field, the light field force it receives is the so-called gradient force. When light passes through a polarizable material, it creates a momentary dipole. These dipoles interact with electromagnetic field gradients, creating a force that points to bright regions of light when the refractive index of these materials is greater than that of the surrounding medium. Conversely, when a substance has a lower index of refraction than the surrounding medium, it experiences a force that pulls it toward darker regions of light. Theories and examples of various biological applications of optical tweezers have been described in detail by many documents such as: "Making light work with optical tweezers, by Block S.M., in Nature, 1992, Volume 360, pages 493-496"; " Forces of a single-beamgradient laser trap on a dielectric sphere in the ray optics regime, by Ashkin, A., in Biophys.J., 1992, Volume 61, pages 569-582"; "Laser trapping in cellbiology, by Wright et al. al., in IEEE J.of Quantum Electronics, 1990, Volume26, pages 2148-2157"; "Laser manipulation of atoms and particles, by ChuS. in Science, 1991, Volume 253, pages 861-866". In order to generate light field radiative force in a chip-based reactor or instrument, a light field and/or light intensity field needs to be generated, for example, by the chip's built-in optical elements and arrays and external light sources or by the chip's built-in Sources of electrical signals for optoelectronic elements and arrays and external structures. In the former case, when the light generated by the optical signal source passes through the built-in optical elements and arrays, the light is processed by these elements/arrays through reflection, focusing, interference, etc., and the light field is generated in the area around the multi-force manipulation chip . In the latter case, when the electrical signal generated by the external electrical signal source is absorbed by the built-in photoelectric elements and arrays, they will generate light, and a light field will be generated around the chip. Other methods can also be used to make the multi-force manipulating chip generate light field, thereby generating light field force.

The micro-device may contain only one functional unit, or may contain multiple functional units for high-throughput analysis, and each functional unit may enable a specific physical force to manipulate the micro-device. For example, the functional unit can be a magnetic substance that can be manipulated by a magnetic field force, an insulating material or a conductive material that can be manipulated by a dielectrophoretic force, a high/low acoustic damping material that can be manipulated by an acoustic field radiation force, a charged material that can be manipulated by an electrostatic force, etc. .

In practical applications, the micro-device has binding substances that can bind or specifically bind to the entity molecules to be separated, detected, and manipulated, and facilitate the manipulation of the micro-device/entity molecule-micro-device complex or enable the micro-device /Entity molecule-microdevice complex manipulation becomes possible unit. The micro-device can also have a group of binders that can bind or specifically bind different entity molecules to be separated, detected, and manipulated, and it is convenient to use different physical forces to manipulate or manipulate the microdevice/entity molecule-microdevice complex. It is a group of units that make it possible to manipulate micro-device/solid molecule-micro-device complexes using different physical forces.

The micro-device may also carry a detectable label or a molecular label. Such detectable labels may be dyes, radioactive and fluorescent substances, and the like. Such molecular tags can be nucleic acids, oligonucleotides, proteins or polypeptides.

In practical application, the microdevice of the present invention can be thin rectangular shape, the ratio of major axis (length) and minor axis (width) is at least 1.2, preferably at least 1.5, the thickness (height) ratio of microdevice is major axis and minor axis. Shafts are small. In another practical application, the micro-device of the present invention comprises at least two rectangular or nearly rectangular strip structures (rod structures) made of paramagnetic substances. The at least two bar-shaped (rod-shaped) structures made of paramagnetic substances are preferably separated and located on both sides of the main axis of the micro-device, separated by a metal film with an optically identifiable coding pattern in the middle. Preferably this metal film contains aluminum. There may be rectangular structures made of different amounts of paramagnetic substances on both sides of the main axis of the micro-device. Microdevices can also have two rectangular structures made of paramagnetic substances along the main axis, and the two rectangular structures made of paramagnetic substances have finger-like protrusions at both ends. It is also possible that the paramagnetic substance forms a rectangular structure along the main axis of the micro-device, and the rectangular structure has finger-like protruding structures at both ends.

Another object of the present invention is to provide a system for forming an array of micro devices.

A system for forming a micro-device array, comprising: 1) a set of micro-device, the micro-device includes: a) a magnetizable base material; b) an optically identifiable coding pattern made on the above-mentioned base material; c) a magnetization axis; and 2) a micro-channel array consisting of a set of micro-channels; the width of the micro-channel is such that a single micro-device can rotate freely in the channel when the micro-device is in an applied magnetic field, A chain of micro-devices cannot be formed when the major axis of the device is perpendicular to the major axis of the microchannel.

In a practical application, the coding portion of the micro-device of the present invention is not realized by platinum, palladium, nickel, cobalt, silver, copper or gold. In another practical application, the constituent materials of the micro-device of the present invention do not include platinum, palladium, nickel, cobalt, silver, copper or gold.

When the micro-device is manipulated in the micro-channel, the micro-device remains in a plane-up or substantially planar-up state, so that it can be optically aligned in a direction perpendicular to the plane in which the length and width of the micro-channel lie. Optical identification code for detection. The height of the micro-channel and/or the adjustment of the constraints on the micro-device by the magnetic field are such that the micro-device does not stand upright in the micro-channel. For example, the height of the microchannel may be no greater than 70% of the length of the major axis of the microdevice.

The micro-channel array device can also include a magnetic field generating device, so as to generate the required magnetic field for manipulating the micro-device to enter and exit the micro-channel, so that the micro-device rotates in the micro-channel. Such a magnetic field generating device can use any suitable device, for example, a permanent magnet, a movable magnet, an electromagnetic unit, a ferromagnetic material, or a micro electromagnetic unit can be used. The generated magnetic field can act on specific positions of the micro-channel array, such as below, in the middle, above and near the micro-channel array.

A third object of the present invention is to provide a method of fabricating an array of micro devices.

There are two methods of fabricating microdevice arrays. A first method of manufacturing an array of micro devices, comprising the steps of:

(1) Provide a group of above-mentioned micro-device, this micro-device comprises a) magnetizable base material; b) the coding pattern that can be made on the above-mentioned base material that can be identified by light; c) a magnetization axis;

(2) Provide a microchannel array that is made up of a group of microchannels; The width of described microchannel is: when described microdevice is in externally applied magnetic field, single microdevice can rotate freely in channel, in described microdevice A chain of microdevices cannot be formed when the main axis is perpendicular to the main axis of the microchannel;

(3) introducing the set of microdevices into the microchannel;

(4) Rotating the micro-device in the micro-channel by an external magnetic field force, and the mutual action of the magnetic field force and the magnetization axis of the micro-device makes the micro-device mutually dispersed.

In a practical application, the coding portion of the micro-device of the present invention is not realized by platinum, palladium, nickel, cobalt, silver, copper or gold. In another practical application, the constituent materials of the micro-device of the present invention do not include platinum, palladium, nickel, cobalt, silver, copper or gold.

When the micro-device is manipulated within the micro-channel, the micro-device remains in a plane-up or substantially planar-up state so as to be optically aligned in a plane perpendicular to the length and width of the micro-channel. The optically recognizable code is detected in the direction. The height of the micro-channel and/or the adjustment of the constraints on the micro-device by the magnetic field should prevent the micro-device from standing upright in the micro-channel. For example, the height of the microchannel may be no greater than 70% of the length of the major axis of the microdevice.

The microdevices can be introduced into the microchannels using any suitable force. For example, microdevices can be introduced into microchannels using magnetic force, fluid force, or a combination of both. There are several ways to introduce microdevices into channels. In a specific implementation, the microdevice may be a microdisk having a certain thickness with two main planes. First place the micro-disk near the micro-channel or at the entrance of the channel, and use a piece of neodymium magnetic material at the outlet of the channel to suck the micro-disk into the channel. The magnetic substance can be rotated to facilitate drawing the microdisc into the channel. In one embodiment of the present invention, the size of the main surface of the microdisk is 90 micrometers by 70 micrometers, and the thickness is about several micrometers. Using the method described above, the microdiscs can be completely filled into five 2 cm long (approximately 120 to 160 micron wide) channels within 3 minutes. The length, width and height of the magnetic strip or rod-like structure in the microdisk are all along the direction of the length, width and height of the microdisk where the magnetic material is located. Since the rate-limiting step is filling the microdiscs along the long axis of the channel, increasing the number of channels does not significantly increase the time required to fill, say, 50,000 microdiscs into 200 2 cm long channels can be completed within 3 minutes. When the direction of the applied magnetic field is perpendicular to the direction of the channel, filling in this way may cause overlapping of "vertically aligned" microdisks. The overlapping of the microdisks can be eliminated by changing the direction of the applied magnetic field from parallel to perpendicular several times. This allows the distance between the "chains" of microdiscs to be increased. When the microdisk is introduced into the channel, it is preferable that the height direction of the microdisk is consistent with the height direction of the microchannel, so that the microdisk can keep the plane upward in the microchannel.

The microdevice may be a microdisk having a certain thickness with two main planes. The microdiscs are filled as described above while maintaining a constant flow rate to increase filling efficiency. When the microdisk is introduced into the channel, it is preferable that the height direction of the microdisk is consistent with the height direction of the microchannel, so that the microdisk can keep the plane upward in the microchannel.

In a specific implementation, the micro-device may be a micro-disk having a certain thickness and having two main planes. The length, width and height of the magnetic strips or magnetic rods in the microdisk are all along the length, width and height of the microdisk where the magnetic material is located. First place the micro-disc close to the micro-channel or the entrance of the channel, use a large neodymium magnetic substance at the exit of the channel to suck the micro-disk into the channel, the direction of the magnetic field generated by this magnetic material and the direction of the channel is vertical. A small neodymium magnet is used above or below the entrance end of the channel, rotated to facilitate the suction of the microdisc into the channel. At the same time, a constant flow rate is maintained to increase the efficiency of filling. The microdisks are filled into the channel according to the "vertical arrangement" mode (that is, the magnetization axis of the microdisks is perpendicular to the direction of the long axis of the channel). This method can reduce the overlapping arrangement of the microdisks in the channel, making the microdisks The alignment of the slices remains uniform. The above-mentioned method of loading and arranging the microdisk will make the magnetic strip structure on or in the microdisk perpendicular to the direction of the long axis of the channel, that is, the magnetic strip structure on or in the microdisk The direction of the long axis of the structure or the rod-like structure is perpendicular or substantially perpendicular to the direction of the long axis of the channel. When the microdisk is introduced into the channel, it is preferable that the height direction of the microdisk is consistent with the height direction of the microchannel, so that the microdisk can keep the plane upward in the microchannel.

The microdevices or microdisks can be introduced into the microchannels at an angle. For example, the micro-device can be filled into the micro-channel with a magnetic field directed in a direction such that the angle between the major axis of the micro-device and the major axis of the micro-channel is no greater than 45 degrees. The orientation of the magnetic field affects the orientation of the microdevice or microdisk and the orientation of the major axis of the microdevice. Usually, when the micro-device can be freely rotated or repositioned, the magnetization axis of the micro-device will be substantially along the direction of the applied magnetic field. For micro-devices or micro-disks, their magnetization axis and the direction of the main axis are consistent, so the direction of the magnetization axis and the direction of the external magnetic field are consistent, that is, the direction of the main axis of the micro-device is consistent with the direction of the external magnetic field. In this way, the micro-device can be filled into the micro-channel with a magnetic field pointing in a direction such that the angle between the main axis of the micro-device and the length direction of the micro-channel is no greater than 45 degrees. Even better, the micro-device can be filled into the micro-channel using a magnetic field directed in a direction such that the angle between the major axis of the micro-device and the length of the micro-channel is no greater than 40, 35, 30, 25 , 20, 15, 10, 5 or 0 degrees.

The method of the present invention further comprises breaking the chain-like arrangement of microdevices prior to or simultaneously with introducing said microdevices into said microchannel. This can be achieved by any suitable means, for example by rotating the direction of the magnetic field between the major and minor axes of the microdevice.

When the microdevices or microdisks are introduced into the microchannels or channels, the microdevices or microdisks remain plane-up. The microdevices or microdisks may be rotated in the channel such that the microdevices or microdisks are spaced apart from each other. The micro-devices or micro-disks are separated from each other, which can be realized through a large-angle rotation, or through multiple small-angle rotations. The rotation angle of the micro-device or micro-disc is at least 45 degrees; preferably, the rotation angle of the micro-device or micro-disk is 90 degrees.

In the specific implementation of the present invention, at least one micro-device can bind one entity molecule, or a group of micro-devices can respectively bind a group of entity molecules. The methods of the invention may be used to perform any suitable manipulation of the entity molecule, such manipulation may be transport, focusing, enrichment, concentration, aggregation, capture, repulsion, suspension, separation, fractionation, isolation, linear or other orientation The movement of the solid molecules on the . The method of the present invention also includes optically detecting the photorecognizable coding pattern on the micro-device to determine the manipulated entity molecule on the micro-device. The determination of the type of the manipulated entity molecule includes obtaining parameters such as the ratio and content of the manipulated entity molecule.

The method of the present invention also includes analyzing the entity molecules on the micro-device by a quantitative method to further obtain quantitative information of the manipulated entity molecules. The quantitative analysis of the manipulated entity molecules includes obtaining parameters such as the quantity and concentration of the manipulated entity molecules. The method of the invention also includes collecting the microdevice bound with the entity molecule through the outlet port. The methods of the present invention also include methods of harvesting physical molecules from collected microdevices.

The second method for manufacturing a micro device array provided by the present invention comprises the following steps:

(1) Provide a group of the above-mentioned micro-devices, the micro-devices include: a) a magnetizable base material; b) an optically identifiable coding pattern fabricated on the above-mentioned base material; c) a magnetization axis.

(2) Make them be located on a plane that is conducive to the rotation of micro-devices;

(3) rotating the micro-devices on the surface by an external magnetic field force, and the micro-devices are dispersed mutually through the combined action of the magnetic field force and the magnetization axis of the micro-devices.

In a certain practical application, the coding portion of the microdevice of the present invention is not realized by platinum, palladium, nickel, cobalt, silver, copper or gold; in another practical application, the composition material of the microdevice of the present invention does not include Platinum, palladium, nickel, cobalt, silver, copper or gold.

The present invention provides a micro-channel array composed of a group of micro-channels. When the above-mentioned micro-device is in an external magnetic field, the micro-channel needs to be wide enough so that a single micro-device can rotate freely in the channel, while the channel It also needs to be narrow enough so that the above-mentioned micro-devices do not form a string of micro-devices when the main axis is perpendicular to the main axis of the micro-channel; c) introducing the above-mentioned group of micro-devices into the above-mentioned micro-channel; d) passing an external magnetic field The force rotates the above-mentioned micro-devices in the micro-channel, and the above-mentioned micro-devices are mutually dispersed through the combined action of the above-mentioned magnetic field force and the preferred magnetizable axes of the above-mentioned micro-devices. In a specific implementation of the invention, the microdevices are introduced into a plane suspended in a solution. The suspension of microdevices can be introduced onto the surface by various methods, such as by adding a sample, or by pumping the suspension into microchannels on the surface. It is also possible to make grooves that are wide first (wider than the size of the micro-device)) and narrower (narrower than the size of the micro-device) on the plane, add the suspension of the micro-device on the plane, and wait until the micro-device is arranged on the plane. The liquid in the suspension in which the microdevices are placed can be removed by various methods, such as suction or pumping.

The present invention can be used on-chip or off-chip to analyze, separate, manipulate or detect any kind of entity molecules that can be processed physically, chemically, biologically, biophysically or Biochemical processing. The entity molecules to be manipulated can be cells, organelles, viruses, molecules, or their complexes. The entity molecule to be manipulated can be a pure substance, or exist in the form of a mixture in which the target entity molecule is only one component of the mixture. For example, cancer cells in the blood of a leukemia patient, cancer cells in a cancer patient's tissue, and fetal cells in a mother's blood can all be used as solid molecules to be isolated, manipulated, or detected. Similarly, different blood cells, such as erythrocytes and leukocytes in blood, can be used as physical molecules to be isolated, manipulated, or detected. DNA molecules, mRNA molecules, specific types of proteins, or the total protein in cell lysates can be used as physical molecules to be isolated, manipulated, or detected.

Cells can serve as physical molecules to be isolated, manipulated or detected. Such cells include, but are not limited to, animal cells, plant cells, fungal cells, bacterial cells, recombinant cells or cells in culture. The animal cells, plant cells, fungal cells and bacterial cells to be manipulated come from a certain genus or subspecies of the kingdom Animalia, Plantae, Fungi and Bacteria respectively. Cells belonging to the ciliates, myxomycetes, flagellates, microspores can also be manipulated. Cells derived from birds such as chickens, vertebrates such as fish and mammals such as rats, mice, rabbits, cats, dogs, pigs, cows, bulls, sheep, goats, horses, monkeys and other great apes and humans can be used as Entity molecules to be isolated, manipulated or detected.

For animal cells, cells derived from specific tissues or organs can be used as entity molecules to be isolated, manipulated or detected. For example, cells of connective, epithelial, muscle or nervous tissue. Similarly, cells in various organs can also be manipulated, such as ocular appendages, annulospiral organs, ear organs, Organs of Cvitz, periventricular organs, Corti's, key organs, enamel, Peripheral organs, female external genitalia, male external genitalia, reproductive organs, Golgi tendon apparatus, gustatory organs, auditory organs, female internal genitalia, male internal genitalia, insertional organs, Jacobsonian organ, neurovascular organs , nerve tendon spindle, olfactory organ, otolithic organ, Rosenmüller organ, sensory organ, olfactory organ, spiral organ, infracommissural organ, infrafornix organ, tactile organ, target organ, gustatory organ, tactile organ, urinary organ, Vestibular organ, vestibulocochlear organ, degenerative organ, visual organ, pear nasal organ, wanderer organ, Weber's organ, and para-aortic body. From animal internal organs such as brain, lung, liver, spleen, bone marrow, thymus, heart, lymph, blood, bone, cartilage, pancreas, kidney, gallbladder, stomach, intestine, testis, ovary, uterus, rectum, nervous system, glands , cells in blood vessels and so on in the body are easier to manipulate. Furthermore, cells from any plant, fungus (such as yeast), bacteria (such as eubacteria or archaea) can be used as the entity molecule to be isolated, manipulated or detected. Recombinant cells from any eukaryotic or prokaryotic organism, such as animals, plants, fungi or bacteria, can be used as entities to be isolated, manipulated or detected. Body fluids from various parts of the body, such as blood, urine, saliva, bone marrow, semen or other ascites cells, and their components such as serum and plasma can be used as solid molecules to be separated, manipulated or detected.

Organelles that can be transported or as conjugates include the nucleus, mitochondria, chloroplasts, ribosomes, endoplasmic reticulum, Golgi apparatus, lysosomes, proteasomes, vesicles, vacuoles or microbodies. Viruses (whether whole virus or any viral structure) that can be transported or as conjugates can be derived from such as the first virus, the second virus, the third virus, the fourth virus, the Type V virus or Type VI virus.

The molecules that can be manipulated, separated or detected can be inorganic molecules such as ions, organic molecules or their complexes. Examples of manipulable ions include, but are not limited to, sodium ions, potassium ions, magnesium ions, calcium ions, chloride ions, iron ions, copper ions, zinc ions, manganese ions, vanadium ions, nickel ions, chromium ions, fluorine ions, silicon ions, tin ions, boron ions or arsenic ions. Examples of organic molecules include, but are not limited to, amino acids, peptides, proteins, nucleosides, nucleotides, oligonucleotides, nucleic acids, vitamins, monosaccharides, oligosaccharides, carbohydrates, lipids, or complexes thereof.

Any amino acid, such as D- and L-amino acids, can be detected, isolated or manipulated using the methods of the invention. In addition, all building blocks of naturally occurring peptides and proteins, including alanine (A), arginine (R), asparagine (N), aspartic acid (D), cysteine (C) , glutamine (Q), glutamic acid (E), glycine (G), histidine (H), isoleucine (I), leucine (L), lysine (K), methionine (M), phenylalanine (F), proline (P), serine (S), threonine (T), tryptophan (W), tyrosine (Y) and valine (V ) can be detected, isolated or manipulated using the methods of the present invention.

Any protein or peptide can be detected, isolated or manipulated using the methods of the invention. For example, membrane proteins (such as receptor proteins), enzymes, transport proteins (such as ion channels and ion pumps), nutritional or storage proteins, contractile or motor proteins (such as actin and myosin), structural proteins on the cell membrane , defensive or regulatory proteins (such as antibodies, hormones and auxins). Both proteinaceous and peptide antigens can be detected, isolated or manipulated using the methods of the invention.

Any nucleic acid, including single-stranded, double-stranded and triple-stranded nucleic acids, can be detected, isolated or manipulated using the methods of the invention. Examples of such nucleic acids include DNA (eg, A-, B-, Z-form DNA) and RNA (eg, mRNA, tRNA and rRNA).

Any nucleoside can be detected, isolated or manipulated using the methods of the invention. Examples of such nucleosides include adenosine, guanosine, cytidine, thymidine, uridine. Any nucleotide can be detected, isolated or manipulated using the methods of the invention, examples of such nucleotides include AMP, GMP, CMP, UMP, ADP, GDP, CDP, UDP, ATP, GTP, CTP, UTP , dAMP, dGMP, dCMP, dTMP, dADP, dGDP, dCDP, dTDP, dATP, dGTP, dCTP and dTTP.

Any vitamin can be detected, isolated or manipulated using the methods of the invention. For example, water-soluble vitamins such as vitamin B1, vitamin B2, niacin, vitamin B3, vitamin B6, vitamin H, folic acid, vitamin B12 and vitamin C can be detected, isolated or manipulated using the methods of the present invention. Similarly, fat-soluble vitamins such as vitamin A, vitamin D, vitamin E and vitamin K can be detected, isolated or manipulated using the methods of the present invention.

Any monosaccharide, whether D- or L-monosaccharide, and whether aldose or ketose, can be detected, isolated or manipulated using the methods of the invention. Examples of monosaccharides include trisaccharides (such as glyceraldehyde), tetrasaccharides (such as erythrose and threose), pentoses (such as ribose, arabinose, xylose, lyxose and ribulose), hexoses (such as Allose, altrose, glucose, mannose, gulose, idose, galactose, talose, and fructose) and heptoses (such as sedum heptulose).

Any lipid can be detected, isolated or manipulated using the methods of the invention. Examples of lipids include triacylglycerides (such as glyceryl stearate, glyceryl palmitate, and glyceryl oleate), paraffins, phosphoglycerides (such as phosphatidylethanolamine, lecithin, phosphatidylserine, phosphatidylinositol, and diphosphatidylglycerol), sphingolipids (such as sphingomyelin, cerebroside, and ganglioside), sterols (such as cholesterol and stigmasterol), and sterol fatty acid esters. Fatty acids can be saturated fatty acids (such as lauryl acid, myristic acid, palmitic acid, stearic acid, arachidic acid, and lignoceric acid) or unsaturated fatty acids (such as palmitoleic acid, oleic acid, linoleic acid , linolenic acid and arachidonic acid).

A fourth object of the present invention is to provide a method for synthesizing a compound library.

There are two methods for synthesizing compound library provided by the present invention. A first method for synthesizing a compound library, comprising the following steps:

1) A set of microdevices is provided, the microdevices include: (a) a base material that can be magnetized; (b) an optically identifiable coding pattern fabricated on the base material; (c) a magnetization axis; The photorecognition coding pattern corresponds to the product of the synthesis reaction on the micro-device;

2) Synthesizing the compound library: the synthesis reaction is realized through a series of synthesis steps; in each step of synthesis, according to the optically identifiable coding pattern on the micro-device, the corresponding synthesis reaction is carried out; after the one-step reaction, according to The coding pattern on the micro-device can be identified by light, and the micro-device is classified, and then the next step of synthesis reaction is performed. Therefore, the product corresponding to the optically recognizable coding pattern is synthesized on the micro-device according to the pre-design.

This method can be used for the synthesis of compound libraries with preset steps

The present invention also provides a compound library synthesized by the above method.

In a certain practical application, the coding portion of the microdevice of the present invention is not realized by platinum, palladium, nickel, cobalt, silver, copper or gold; in another practical application, the composition material of the microdevice of the present invention does not include Platinum, palladium, nickel, cobalt, silver, copper or gold.

The second method for synthesizing a compound library provided by the present invention comprises the following steps:

1) A set of microdevices is provided, the microdevices include: (a) a base material that can be magnetized; (b) an optically identifiable coding pattern fabricated on the base material; (c) a magnetization axis; The photorecognition coding pattern corresponds to the product of the synthesis reaction on the micro-device;

2) Synthesizing the compound library: the synthesis reaction is realized through a series of synthesis steps; after each step of synthesis, according to the optically identifiable coding pattern on the micro-device, the corresponding synthesis reaction is recorded, and then the next step of the synthesis reaction is carried out . Thus, the corresponding relationship between the photorecognizable coding pattern on the micro-device and the synthesized product on the micro-device is obtained.

This method can be used for the synthesis of random compound libraries.

The present invention also provides a compound library synthesized by the above method.

In a certain practical application, the coding portion of the microdevice of the present invention is not realized by platinum, palladium, nickel, cobalt, silver, copper or gold; in another practical application, the composition material of the microdevice of the present invention does not include Platinum, palladium, nickel, cobalt, silver, copper or gold.

The microdevices can be sorted using any suitable method. For example, sorting can be performed using a microchannel array consisting of a set of microchannels. The micro-channel is wide enough so that a single micro-device can rotate freely in the channel, and the channel is narrow enough so that the above-mentioned micro-device will not form a micro-device string under the condition that the main axis of the micro-device is perpendicular to the main axis of the micro-channel. Through the joint action of the external magnetic field force and the magnetization axes of the above-mentioned micro-devices, the above-mentioned micro-devices are mutually dispersed. The height of the micro-channel and/or the limitation of the micro-device by the magnetic field prevent the micro-device from standing upright in the micro-channel. The height of the micro-channel is not greater than 70% of the length of the major axis of the micro-device. After the micro-devices are arranged in the micro-channel, the optical analysis method can be used to determine the optically recognizable coding pattern on the individual micro-devices. Microdevices can be manipulated in the same way as individual microdevices and sorted into different regions/locations/reaction chambers based on the optically identifiable coded patterns carried on the microdevices. For example, microelectromagnetic tips can be used for sorting. The micro-electromagnetic tip can generate a magnetic field at the tip to absorb a specific micro-device from a channel lined with micro-device (in this case, the upper part of the channel must be open), move and deliver/distribute a single micro-device to different area/position, reaction chamber. In a specific implementation of the present invention, the micro-device moves out of the micro-channel under the combined action of magnetic force and fluid force, and at the outlet end of the channel, the micro-device can be transported to different places under the combined action of magnetic force and fluid force. Area.

The above sorting can also use magnetic force to specifically capture the required micro-devices after each step of the reaction and send them into a suitable reaction chamber. The arrangement cavity of the micro-device can be fabricated on the upper surface or the lower surface of the arranged micro-device by using photoresist. When irradiated by light of a specific wavelength, the photoresist in the irradiated area can be dissolved and washed away, and the micro-devices that were just covered are exposed and can be removed by magnetic force. A programmable digital micromirror array (Singh-Gasson et al. Maskless fabrication of light-directed oligonucleotides microarrays using a digital micromirror array. Nature Biotechnology, 17: 974-978 (1999)) or other similar maskless Modular array combiner to direct the light source.

Sorting using magnetic force can also be achieved through sorting channels. Micro-devices with magnetization axes can be aligned in a magnetic field and separated from each other under the action of the magnetic field force. Under the push of the liquid, they are "vertically arranged" (the magnetization axis is perpendicular to the long axis of the channel) ) in the form of filling into the channel. By presetting an appropriate liquid-liquid (immiscible liquid, such as hexane and water) or gas-liquid interface, the surface tension of the solution can be increased to limit the movement of the micro-device in the solution. In the specific implementation of the present invention, the channel for introducing the micro-device and the channel for sorting are separated by a micro-valve (the design, manufacture and use of valves are common knowledge in the related field). An air bubble can be introduced into the channel by making a hole in a suitable position near the outlet end of the channel introducing the microdevice, this bubble being located just between the last and the penultimate microdevice. Then the valve is opened, and only the last micro-device can be introduced into the sorting channel by magnetic force, while other micro-device are blocked behind the introduced air bubbles. Then close the valve, and use magnetic field force, fluid force or other appropriate physical force (such as dielectrophoretic force) to sort the micro-device in the sorting channel into an appropriate reaction chamber. Then use fluid force (such as pumping liquid) to discharge the air bubbles introduced into the channel end of the micro-device out of the hole at the outlet end of the channel, so that the position of the micro-device in the channel moves forward to the outlet end, and then repeat the above-mentioned sorting operation . The holes for introducing and evacuating air bubbles must be considerably smaller than the size of the micro-device. A similar and faster sorting solution is to introduce a bubble between every pair of micro-devices in the channel, adjust the appropriate magnetic force and fluid force, so that the micro-devices pass through the channel in a segmented flow. This method is similar to the segmented flow method widely used by Technicon (see US Pat. Nos. 2,797,149 and 3,109,713). When using the segmented flow method, both the magnetic field force and the fluid force are important influencing factors. In addition to this, the third influencing factor is the surface tension of the liquid. The surface tension of the liquid can be determined by adding a solvent or Additives (such as detergents) for adjustment. The adjustment of the surface tension by adding a solvent employs a common technique in the related art.

Microdevices can also be sorted using devices capable of manipulating particles, such as particle switches. As shown in Figure 7, the microchannel array device can include an area in which the microdevices can be manipulated, a sample loading zone for adding samples of the microdevices to the device, and outlet channels for discharging the microdevices, collection zone Combined part with the fluid pipeline. The US patent application "Apparatus for switching and manipulating particles and methods of use thereof" (application number: No.09/678,263, date of submission on October 3, 2000) provides a device that can be used to manipulate and sort particles. These devices and devices and their methods of use can be applied to the sorting of micro devices of the present invention. For example, sorting can be performed on particle switching devices by traveling wave dielectrophoresis. The particle switch device has at least three electrode groups independently loaded with electrical signals. There are units with multiple branch structures on the particle switching device. These branch structures have a common connection area. When the three or more electrode groups are loaded with different phase electrical signals, the particles are subjected to different traveling wave dielectrophoresis. The force can move along the branches of the device, from one branch to another branch through the common connection area of each branch. The ends of these branches (opposite the common connecting region of the branches) can be used as inlets or outlets. In a specific implementation of the present invention, the particle switch device has 3 inlets (outlets). One of the ports can be used as an inlet, and the other two ports can be used as outlets. The micro device of the present invention can be introduced into the particle switch device through the inlet, transported along the branch under the action of the electric signal, and then output the device through one of the outlets. The optically identifiable code pattern on the micro-device can be optically detected in the particle switch, and then according to the photo-recognizable code pattern carried on the micro-device, an appropriate electrical signal is applied to the electrode of the micro-particle switch device, so that the micro-device is activated Sorting to output devices from one outlet. An array device composed of such particle switching devices can sort microdevices to more than two outlets. A description of such an array of particle sorting devices can be found in US Patent Application No. 09/678,263.

Microdevices can also be sorted by using a fluidic system with one inlet and multiple outlets. A fluidic system can transport microdevices from an inlet to any outlet. Each micro-device will flow through the optical decoder in the fluid system. The decoder can recognize the optically recognizable coding pattern on the micro-device, and then transport the micro-device to different outlets by changing the type of fluid according to the code.

Other methods of sorting based on optically identifiable coded patterns on the microdevices may also be used.

Any number of compounds can be synthesized on one such microdevice, for example, one or a group of compounds can be synthesized on one microdevice. Preferably, only one compound is synthesized on one microdevice.

The methods of the invention can be used to synthesize any kind of library. The components of the library can be polypeptides, proteins, oligonucleotides, nucleic acids, vitamins, oligosaccharides, carbohydrates, lipids, small molecules or their aggregates and complexes. The synthetic compound library includes a group of entity molecules related to a certain biological pathway. This group of entity molecules has the same or similar biological functions, and they are all expressed in the same cell cycle, expressed in the same cell type, and expressed in the same cell type. Expressed in a tissue type, expressed in the same organ, expressed in the same developmental stage. The expression or activity of the entity molecule may be changed by the disease, or by the type and stage of the disease, or by drugs or other treatments.

A synthetic library can be a nucleic acid library, containing a variety of nucleic acid molecules, such as fragments of DNA or RNA molecules covering a genome (eg, the human genome). Preferably each nucleic acid fragment in the library comprises at least 10, 15, 20, 25, 50, 75, 100, 200 or 500 nucleotides.

The synthetic library can also be a protein/polypeptide library, containing a variety of protein/polypeptide molecules, such as protein/polypeptide fragments covering all protein or polypeptide sequences (such as human protein or polypeptide sequences) in the coding organism. Preferably each protein/polypeptide fragment in the library comprises at least 10, 15, 20, 25, 50, 75, 100, 200 or 500 amino acid residues.

Compound libraries can be synthesized using the methods described above.

The invention can be widely used in the fields of separation, detection, manipulation and compound library synthesis of entity molecules/molecules.

Description of drawings

FIG. 1 is a schematic diagram of the structure of a micro device (micro disk).

FIG. 2 shows a possible arrangement of a plurality of microdisks confined on a plane under the action of a magnetic field in the direction of the arrow in the figure.

Fig. 3 is a short chain arrangement structure in which a plurality of microdisks are confined on a plane and then introduced into a channel under the action of a magnetic field in the direction of the arrow in the figure.

FIG. 4 shows the arrangement of the short chains of microdisks shown in FIG. 3 after the direction of the magnetic field is rotated by 90 degrees as indicated by the arrow.

Figure 5 is an example of a minidisk with different magnet bar types.

Figure 6 shows two different coding patterns.

Fig. 7 shows a microchannel device composed of a sample loading area, a microchannel, a collection area and a combination part of a fluid pipeline.

Figure 8 is an example of 4 minidiscs.

FIG. 9 shows that under the action of an external magnetic field as shown by the arrows in the figure, the microdisks form a linear chain arrangement on the glass surface.

FIG. 10 shows that under the action of an external magnetic field as shown by the arrows in the figure, the microdisks form a linear chain arrangement with branches on the glass surface.

Figure 11 shows that the microdisk is confined in a channel with a width of 130 micrometers under the action of an applied magnetic field as indicated by the arrows in the figure.

Detailed ways

definition

Unless otherwise defined, all scientific and technical terms used in the present invention are to be understood according to the general definitions in the field to which they belong. In general, the nomenclature herein and the fabrication and experimental procedures described below are common and frequently cited in this technical field. These procedures all use conventional methods, as well as the technique and the general literature. Directional terms such as "upper", "lower", "upper", "lower" and similar terms refer to the orientation of parts of the device when in use. The nomenclature in this document is generic and is commonly used in this technical field. Where there are differences between the terms and definitions and the unified terms and definitions in the literature, the definitions in this article should be used. The following terms in the present invention should be understood according to the following explanations if there is no other explanation.

"Magnetic substance/material" refers to a substance that is magnetic and can be affected by a magnetic force.

"Magnetizable substances/materials" are those substances that can be subjected to a magnetic field, and when placed in a magnetic field, can be magnetized to induce magnetic dipoles. Magnetizable substances include, but are not limited to, paramagnetic, ferromagnetic or ferrimagnetic substances.

"Paramagnetic substance/material" means a substance having the property that its atoms, iron or molecules have permanent magnetic dipoles. Due to the thermal motion of the atoms or molecules in the material, the directions of these magnetic dipoles are randomly pointing in various directions, so the material is generally not magnetic. However, in the external magnetic field, the magnetic dipoles pointing in all directions will point to the direction parallel to the magnetic field under the action of the magnetic field, because the energy of these magnetic dipoles pointing to the direction parallel to the magnetic field is higher than the energy not pointing to the parallel direction. The lower the value, the more stable it is. In this way, a net magnetism is produced in the direction parallel to the magnetic field, which also increases the magnetic susceptibility of the material. Further information on "paramagnetic substances" can be found in many related documents, such as Page 169-page 171, Chapter 6, in "Electricity and Magnetism" by B.I Bleaney and B.Bleaney, Oxford, 1975.

A "ferromagnetic substance/material" has a large (positive) magnetic susceptibility coefficient, which is related to the strength of the applied magnetic field. In addition, when the external magnetic field is removed, the ferromagnetic substance still retains a part of its magnetism, which is called "remanent magnetism". Further information on "ferromagnetic substances" can be found in many related documents, such as Page 171-page 174, Chapter 6, in "Electricity and Magnetism" by B.I Bleaney and B.Bleaney, Oxford, 1975.

A "ferrimagnetic substance/material" is similar to a ferromagnetic substance in that it has both spontaneous and remanent magnetism. However, the spontaneous magnetic dipoles of this material do not reach the value obtained by calculating that all the magnetic dipoles are arranged in one direction, so it is called "ferrimagnetic material". For further information on "pressurized magnetic substances", you can refer to many related documents, such as Page 519-524, Chapter 16, in "Electricity and Magnetism" by B.I Bleaney and B.Bleaney, Oxford, 1975.

"Optically identifiable coding pattern" refers to any coding pattern that can be identified and analyzed by optical methods. Any light-recognizable property can be used as a feature of the coded pattern. For example, the optically recognizable pattern can be given by the composition of the substance itself: the holes punched on the base material are arranged in a certain form; other materials with different optical properties from the base are fixed on the base material in a certain way. The encoding method of the optically identifiable coding pattern can be based on the substrate itself or the hole punched on the substrate or the figure, number, position distribution, optical property, material composition or a combination of the above-mentioned methods fixed or placed on the substrate material. In order to facilitate the optical analysis of the optically coded pattern, the micro-devices preferably have markings for positioning. For example, for disc-shaped micro-devices, when the micro-devices are placed flat, it is difficult to distinguish the front or back of the micro-devices, which will cause difficulties in identification. And just can determine which side is the front side that has coding pattern by positioning mark. In the present invention, a one-dimensional or two-dimensional barcode can be used as the optically recognizable coding pattern.

"The optically identifiable coding pattern made on the base material" means that the optical coding pattern is located on the base material or made in the base material, as long as it is convenient for optical detection of the code. For example, the optically recognizable coding pattern can be located on or on the surface of the substrate, or can be embedded in the substrate. If the substrate is composed of multiple layers of materials, the optically recognizable coding pattern can be located on or on the surface layer, or can be fabricated on one or more layers of materials.

"Making an optically identifiable coding pattern on a base material by micromachining technology" refers to fabricating an optically identifiable coding pattern on a base material by micromachining technology. Various semiconductor processing techniques can be used, such as photolithography, wet etching, masking, reactive ion etching, and deep reactive ion etching, among others.

"Main axis of a micro-device" refers to the longest dimension among the three-dimensional dimensions of a micro-device. If the micro-device is a thin disk-shaped structure, the height of the micro-device is the thickness of the micro-device. In such thin disc-shaped microdevices, the major axis is the diameter of any circle parallel to the disc plane of the disc. In a practical application of such a disc-shaped micro-device, the optically identifiable coding pattern is located on a plane parallel to the disc surface, which may be located on the surface of the disc or in the area between the upper and lower discs. If the micro-device is a thin rectangular structure, first define the three dimensions of the device, which are the major axis (eg length), the minor axis (eg width) and the height (thickness of the rectangular disk). In this case, the major axis of the micro-device should be larger than the minor axis and height. The minor axis should also be equal to or greater than the thickness of the microdevice. In addition to the above-mentioned shapes of the micro-device, the disk surface can also have other shapes.

"The micro-device has a magnetization axis" means that when the micro-device is placed in a magnetic field, according to the difference between the direction of the magnetic field and the angle between the axes of the micro-device when placed, the micro-device is affected by the magnetic field differently, Assume that the micro-device is placed in a medium with low frictional resistance (no frictional resistance or low frictional resistance), or is placed in a medium with low frictional resistance (no frictional resistance or low frictional resistance). On the plane, the micro-device will be rotated or positioned at a certain position so that it is in the lowest energy state or stable state in the applied magnetic field. When micro-devices are placed in a medium with low frictional resistance (no frictional resistance or small frictional resistance), or placed in a medium with small frictional resistance (no frictional resistance or small frictional resistance) On the plane, and in such a state of the lowest energy, the axis of the micro-device corresponding to the direction of the magnetic field is called the magnetization axis of the micro-device. The magnetization axis is determined by the geometry of the microdevice, eg, the ratio of the major and minor axes of the microdevice, or the compositional and structural parameters of the microdevice. Depending on the geometry of the micro-device, the magnetization axis can be a single axis in one direction, multiple axes in multiple directions, or any axis in a plane. When the micro-device in the magnetic field is induced to be magnetized and reaches the lowest energy state, the induced magnetization (its absolute value) generated in the direction of the magnetization axis is greater than or equal to the induced magnetization generated in other axis directions. The micro-device described in the present invention will produce induced magnetization phenomenon in the external magnetic field, and the interaction between the induced magnetic moment and the external magnetic field makes the micro-device rotate in the magnetic field or locate at a certain position in the magnetic field. The induced magnetization in the microdevice along the direction of the magnetization axis (its absolute value) should be at least 20% greater than the induced magnetization in the direction of at least one other axis. More preferably, the induced magnetization (its absolute value) along the direction of the magnetization axis in the microdevice should be at least 50%, 70% and 90% greater than the induced magnetization along at least one other axis, respectively. Preferably, the induced magnetization (its absolute value) along the direction of the magnetization axis in the microdevice should be at least 1, 2, 5 times larger than the induced magnetization generated in the direction of at least one other axis 10 times, 20 times, 50 times or even hundreds of times. When a micro-device is placed in an external magnetic field, it is a dynamic process for a micro-device to go from an initial state to a state with the lowest energy or a stable state, and it takes a certain amount of time. In the presence of friction or other forces such as gravity, the magnetization axis and the direction of the applied magnetic field may not exactly coincide when the microdevice reaches a steady state. Many factors, such as the geometry of the micro-device, the direction and strength of the applied magnetic field and others (such as the friction force generated by the surface of the support medium when the micro-device is on the surface of the support medium can be such a factor) will affect the micro-devices. magnetization axis. By changing and adjusting these factors, the magnetization axis can be basically along the direction of the applied magnetic field when the micro-device reaches a stable state. For example, a thin disk-shaped micro-device containing a thin disk-shaped magnetizable material. The magnetization axis of the micro-device should lie in a plane parallel to the surface of the micro-device disc (ie, the disc surface of the magnetizable material in the disc). When such a microdevice is placed in a magnetic field, even though the thin disk-shaped microdevice is initially in a plane perpendicular to the direction of the applied magnetic field, the microdevice rearranges so that the plane of the sheet-shaped microdevice is parallel to the direction of the applied magnetic field Or basically parallel. If the micro-device is a thin rectangular structure, the length, width and height of the magnetic structural unit (such as a magnetic rectangular rod) composed of magnetizable materials are consistent with the orientations of the length, width and height of the micro-device respectively. The direction of the magnetization axis of the micro-device with such a structure and the direction of the length of the micro-device are consistent with the direction of the length of the magnetizable material in the micro-device.

"The magnetization axis may be substantially along the direction of the applied magnetic field" means that the included angle between the direction of the magnetization axis and the direction of the applied magnetic field is less than or equal to 45 degrees. In a better situation, the angle between the direction of the magnetization axis and the direction of the applied magnetic field is less than or equal to 15 degrees; in the best situation, the direction of the magnetization axis is completely along the direction of the applied magnetic field. When the magnetization axis of a micro device is its main axis, "the magnetization axis may be substantially along the direction of the applied magnetic field" means that the included angle between the direction of the main axis and the direction of the applied magnetic field is less than or equal to 45 degrees. For a thin rectangular structure, the magnetization axis is the main axis of the micro-device, and under the action of an external magnetic field, the micro-device can be arranged in a chain along the main axis. When the direction of the applied magnetic field is rotated by more than 45 degrees (for example, 90 degrees), the micro-devices also rotate by the same or similar degree, so that the micro-devices in the chain micro-devices are separated from each other.

"The magnetization axis of the micro-device may be substantially along the direction of the main axis of the micro-device" means that the included angle between the direction of the magnetization axis of the micro-device and the main axis of the micro-device is less than or equal to 45 degrees. In a better case, the angle between the direction of the magnetization axis of the micro-device and the main axis of the micro-device is less than or equal to 15 degrees; in the best case, the direction of the magnetization axis of the micro-device is completely along the direction of the main axis of the micro-device .

"The microdevices arranged in a chain are substantially separated from each other" means that the microdevices are substantially separated from each other so that each microdevices can be identified by reading the optically recognizable coding pattern carried on the microdevices. microdevices or perform analysis on microdevices. The degree to which individual microdevices are separated is determined by a number of factors, such as the type of microdevices, number, distribution of the photorecognizable coding pattern, geometry of the microdevices, method of reading the photorecognizable coding pattern, and identification or identification of the microdevices. is the purpose of the analysis, etc. As long as it does not affect the reading of the optically identifiable coding pattern carried on the micro-devices to determine the micro-devices or analyze the micro-devices, the possible mutual contact or partial overlap between the micro-devices is acceptable. In some cases, it is desirable that the micro-devices be completely separated without any contact or overlap.

"The width of the micro-channel is such that when the micro-device is in an external magnetic field, a single micro-device can freely rotate in the channel, and a micro-device chain cannot be formed when the main axis of the micro-device is perpendicular to the main axis of the micro-channel." It means that the width of the micro-channel is equal to or greater than the longest dimension of the micro-device. For example, when the micro-device is a rectangular structure, the width of the micro-channel is taken as the size of the diagonal of the rectangle, so that the micro-device can rotate freely in the channel. . At the same time, the width of the channel cannot be greater than 150% of the longest dimension of the micro-device. For example, when the micro-device has a rectangular structure, the width of the micro-channel is taken as the size of the diagonal of the rectangle. In this way, if the main axis of the micro-device is substantially perpendicular to the main axis of the micro-channel, the micro-channel can prevent the micro-device in the channel from being arranged in a chain (a chain containing at least two micro-device). In the best case, the width of the channel should not be greater than 140%, 130%, 120%, 110%, 105%, or 102% of the longest dimension of the microdevice. "A micro-device chain cannot be formed when the major axis of the micro-device is perpendicular to the major axis of the microchannel." means that after rotation, the micro-devices arranged in a chain are substantially separated from each other. The width of each microchannel in the microchannel array can be the same, but this is not required. Each microchannel in the array of microchannels is such that a microdevice can be rotated within the channel. Here, the major axis of the microchannel is the direction along the length of the microchannel.

"The main axis of the micro-device is substantially perpendicular to the main axis of the micro-channel" means that the included angle between the main axis of the micro-device and the main axis of the micro-channel with the micro-device is greater than or equal to 45 degrees. More preferably, the included angle between the major axis of the micro-device and the major axis of the micro-channel with the micro-device is greater than or equal to 50, 55, 60, 65, 70, 75, 80, 85 and 90 degrees. Here, the major axis of the microchannel is the direction along the length of the microchannel.

"The height of the micro-channel and/or the limitation of the micro-device by the magnetic field prevents the micro-device from standing up in the micro-channel" refers to limiting the height of the micro-channel or the limitation of the micro-device only by the magnetic field, Or the sum of the above two is enough to prevent the micro-device from the situation that the main axis and the height direction of the micro-channel are substantially parallel in the micro-channel. The dimensions of the microchannel include length, width and height. The length of the microchannel corresponds to its major axis. The height of the microchannel is along a direction perpendicular to the plane in which the microchannel is located. The width of the microchannel is the third dimension of the microchannel. "The main axis and the height direction of the microchannel are substantially parallel" means that the included angle between the main axis of the micro device and the height direction of the microchannel is less than or equal to 45 degrees. If the above arrangement of micro devices can be prevented only by using the method of magnetic field confinement, the height of the micro channel itself can be ignored.

"The optically identifiable coding pattern corresponds to the product to be synthesized on the microdevice" means that the compound synthesized on the microdevice is predetermined according to the optically identifiable coding pattern on the microdevice. The code determines how the compound is synthesized on the tiny device. For example, the code may include a plurality of numbers, and each number represents a specific synthetic step. Such a group of numbers in the code represents a synthetic step to synthesize the desired product. Synthesis can also be routed from the entire code rather than a portion of the code.

"According to the optically identifiable code, the micro-devices are re-sorted after each step of the synthesis reaction" means that the synthesis steps of the compounds on each micro-device are pre-marked by the optically identifiable code on it, and after the previous synthesis reaction is completed, According to the coding on the micro-devices, the micro-devices are sorted, and then the next reaction is carried out.

"Conductive or dielectrically polarizable substance" refers to any substance that can be subjected to dielectrophoretic forces under specified conditions. According to the electrical and dielectric properties of the substance itself, under certain conditions, the substance can be subjected to positive or negative dielectrophoretic force. Such specific conditions include, but are not limited to, an applied electric field of a specific frequency, a medium with certain electrical and dielectric properties.

"Optical marker substances" refer to those optically visible substances that can be used to label the entity molecules to be transported. Quantum dots are one such optical marker substance.

"Scattered light detectable microparticle" refers to a microparticle that, when illuminated with light under specified conditions, produces a unique and resolvable scattered light. Nano-sized particles with the property of "resonance light scattering (RLS)" are one such "scattered light detectable particles".

"Quantum dots" are a class of fluorescent markers composed of water-soluble semiconductor nanocrystals. A unique property of quantum dots is that their fluorescence spectrum is determined by the diameter of the nanocrystals. "Water-soluble" herein means that the substance has sufficient solubility in aqueous solutions (water or solutions made of water or solutions of physiological conditions) or can be suspended in these solutions. Information on quantum dots can be found further in the literature. Generally, quantum dots with relatively good monodispersity can be prepared, for example, the diameter deviation of the inner core of the prepared quantum dots is within 10%.

"Chip" refers to a surface on which at least one processing or manipulation can be performed. For example: transfer, separation, aggregation, enrichment, concentration, physical fragmentation, mixing, combining, analysis, etc. Chips can be made from solids or semisolids, and the materials can be porous or dense. A particular process may be performed thereon, such as a physical, chemical, biological, biophysical, or biochemical process. Microfabricated structures such as grooves, channels, electrode units, and piezoelectric sensors are fabricated or integrated on the chip, on which physical, biophysical, biological, and chemical reactions or treatments can be performed. A chip is a thin sheet. The requirement on the surface area of the chip is not strict, for example, it can be from 1mm ² to 0.25m ² . Preferably the chip used has a surface area of from 4 mm ² to about 25 cm ² . Chips can have different shapes, regular shapes such as rectangles, circles, ovals or other irregular shapes. Chip surfaces can be flat or uneven. A chip with an uneven surface should include grooves, cavities and other structures on the surface obtained by processing or etching on the chip surface.

"The method of generating physical fields on the chip" refers to any substance, structural unit or combination thereof that can be generated on the chip, or cooperate with the structural units built in the chip to generate the required physical field.

"Physical field", other terms include "physical field within a certain spatial range" or "physical field generated within a certain spatial range" refers to a spatial range with the following characteristics. When an entity molecule with appropriate properties is placed into this space (that is, into the physical field), the entity molecule is subjected to a force as a result of the interaction between the entity molecule and the field. Entity molecules are manipulated in the field by the forces exerted on them by the field. Typical fields include electric, magnetic, acoustic, light, and velocity fields. In the present invention, the physical field always exists in the medium within a certain space range, and these manipulated entity molecules are usually suspended, or dissolved, or more generally placed in the medium. Typical media are liquids such as water or non-aqueous liquids, or gases. Depending on the nature of the field, the electric field can exert an electrophoretic force on the charged entity molecule, or a conventional dielectrophoretic force and/or a traveling wave dielectrophoretic force on the charged or neutral entity molecule. The magnetic field can exert a magnetic field force on the magnetic entity molecule; the sound field can exert an acoustic field radiation force on the entity molecule; and the light field can exert an optical field radiation force on the entity molecule. The velocity field in the medium within a certain spatial range refers to the velocity distribution of the medium moving within this spatial range. Various mechanisms can cause the movement of the medium and the medium at different locations exhibits different velocities, thus creating a velocity field. Velocity fields can exert mechanical forces on solid molecules.

"Medium" refers to a fluid carrier, such as a liquid or a gas. There are solid molecules suspended, contained or dissolved therein or solid molecules connected to micro-devices.

"Microfluidic application" refers to the use of tiny devices, whose basic structural units are on the submicron to centimeter scale, for fluid manipulation, processing and execution of special biological, biochemical or chemical reactions. These particular areas include, but are not limited to, biochips (such as microchips for biologically related reactions and manipulations), chemical chips (microchips for chemical reactions), or hybrid chips. The characteristic size of the basic unit in the device refers to the size of a certain dimension in the device. For example, the characteristic dimension of a unit of a circular structure (eg, a circular electrode plate) refers to the diameter of the circular structure. If the structural element is a thin rectangular line, its characteristic structure is the length or width of the line.

"Built-in structural units on the substrate" means that these structural units are embedded in the substrate or that these structural units are located on the substrate and connected to the substrate. In practical applications, such built-in structural units are directly processed on the substrate. For example, spiral electrodes are microfabricated on glass substrates as described in "Dielectrophoretic manipulation of cells using spiral electrodes by Wang et al., Biophys. J., 72: 1887-1899 (1997)". Here, the helical electrode is a "built-in" structural unit on a glass substrate. In practical applications, the built-in structure can be processed on a substrate first, and then the substrate containing the built-in structure can be connected with another substrate. This "built-in" structure is thus present on both substrates. In practical applications, this built-in structure can be directly connected to one side of the substrate. For example, thin electrical wires can be used as electrodes to generate an electric field. These electrodes can be attached directly to the glass substrate. In this example, the electrical leads are "built-in" structures on a glass substrate. In the present invention, the described built-in structure capable of generating physical force and/or physical field on the chip or substrate can be connected to an external signal source.

The characteristic size of the "microstructure" is between 1 micrometer and 20 millimeters, and the size of the internal structure on the device to generate the required physical force should be coordinated with the requirements of the microfluidic application.

"Entity molecule" refers to any substance that can be analyzed, separated, manipulated, measured and detected on the micro-device of the present invention. In general, the characteristic dimensions of these substances should not exceed 1 cm. For example, if the substance is spherical or nearly spherical, its characteristic dimension refers to the diameter of the spherical or nearly spherical shape. If the substance is a cube or near cube, its characteristic dimensions are the side lengths of the cube or near cube. If the substance has an irregular shape, its characteristic size is the average of the lengths of its largest and smallest axes. Entity molecules can be, but are not limited to, cells, organelles, viruses, particles, molecules (such as proteins, DNA and RNA), or aggregates or complexes thereof.

Entity molecules that can be analyzed, separated, manipulated, measured and detected can include - solid (eg: glass beads, latex particles, magnetic beads), liquid (eg: liquid droplets) or gaseous particles (eg: bubbles), dissolved Particles (such as molecules, proteins, antibodies, antigens, lipids, DNA, RNA, molecular complexes), suspended particles (such as glass beads, latex particles, polystyrene beads), particle molecular complexes (such as: DNA molecular magnetic bead complexes formed by immobilizing DNA molecules on the surface of beads, or protein-polystyrene bead complexes formed by coating polystyrene beads with protein molecules). Particles can be organic (eg mammalian cells or other cells, bacteria, viruses or other microorganisms) or inorganic (eg metal particles). Microparticles can have different shapes (eg, spherical, ellipsoidal, cubic, discus, needle-like) and different sizes (eg, nanoscale gold spheres, micron-scale cells, millimeter-scale particulate polymers). Particles include, but are not limited to, biomolecules (such as DNA, RNA, chromosomes), protein molecules (such as antibodies), cells, colloidal particles (such as polystyrene beads, magnetic beads) and other biomolecules (such as enzymes, antigens, , hormones, etc.).

"Plant" means any photosynthetic eukaryotic multicellular organism in the genus Planta, characterized by the ability to produce embryos, containing chloroplasts, having cell walls of cellulose composition, and being immobile.

"Animal" means any multicellular organism in the genus Animals, characterized by the ability to move, lack of a photosynthetic mechanism, marked response to stimuli, limited development, and generally fixed body structure. Examples of animals include, but are not limited to, birds such as chickens, vertebrates such as fish and mammals such as rats, mice, rabbits, cats, dogs, pigs, cows, bulls, sheep, goats, horses , monkeys and other great apes), etc.

"Bacteria" refers to small prokaryotes (characteristic size on the order of 1 micron) with ribosomes and closed circular DNA with a sedimentation coefficient of about 70s. Bacterial proteins are synthesized differently than eukaryotes. Many antibiotics inhibit bacterial protein synthesis without affecting the infected host.

"Eubacteria" refers to another large subclass of bacteria besides archaea. The vast majority of Gram-positive bacteria, cyanobacteria, mycoplasmas, enterobacteria, pseudomonas, and chloroplasts are eubacteria. The plasma membrane of eubacteria contains ester-linked lipids and, if present, the cell wall, which contains peptidoglycan; no endogens are found in the genomes of eubacteria.

"Archaebacteria" refers to another large subclass of bacteria besides eubacteria. There are three main groups of archaea: halophiles, methanogens, and an extremely thermophilic fungus that depends on sulfur for survival. Archaea are distinguished from eubacteria mainly by differences in ribosome structure and cell membrane composition, and (in some cases) archaeal genomes have introns.

A "virus" is an organism that lives inside a host cell without the structural characteristics of a cell, having a coat of DNA or RNA and proteins. Viruses range in size from about 20 to about 300 nanometers. The genome of the first type of virus (Baltimore classification) is double-stranded DNA; the genome of the second type of virus is single-stranded DNA; the genome of the third type of virus is double-stranded RNA; the genome of the fourth type of virus is single-stranded positive-strand RNA , the genome itself is the mRNA of the virus; the genome of the fifth type virus is a single-stranded anti-strand RNA, and the genome is a template for synthesizing mRNA; the genome of the sixth type virus is a single-stranded positive-strand RNA, but in the replication and mRNA synthesis Both use DNA as the medium. Most viruses are identified by the diseases they cause in plants, animals and prokaryotes. Prokaryotic viruses are called bacteriophages.

"Fungi" are a group of eukaryotic organisms that grow in irregular aggregates without roots, stems or leaves, and lack chlorophyll or other photosynthetic pigments. Each fungus (plankton) is a unicellular filament with branched hyphae, surrounded by a cell wall (containing glucan and chitin or both), containing a true nucleus.

"Conjugate" refers to any substance that can be linked to an entity molecule with a certain affinity and specificity. The conjugate can be, but not limited to, cells, organelles, viruses, particles, aggregates or complexes, or aggregates or complexes of molecules, and can also be special molecules, such as antibodies, single-stranded DNA, etc. The combination can also be the surface-coated substance of the micro-device in the present invention. Conjugates can also be part of microdevices. Among the constituent materials of the micro-device of the present invention, in addition to being the substrate of the micro-device itself, it can also have affinity binding activity for some entity molecules, and can directly function as a conjugate.

"A substance that facilitates/enables the manipulation of a microdevice or a solid molecule/microdevice complex" refers to a substance that is itself manipulable and allows the solid molecule/microdevice complex to be manipulated by physical forces. Such substances can be, but are not limited to, molecules, cells, organelles, viruses, particles, or complexes of the above substances, for example. Such substances can also be, but are not limited to, substances with special physical and chemical properties produced by deposition or other processes. Such substances can be metal films made of metals such as gold, chromium, titanium or platinum, which can not only be used as constituent materials of micro-devices, but also can increase the conductivity of micro-devices. Such substances can also be insulating substances such as plastic polymers such as polystyrene, which can not only be used as constituent materials of micro-devices, but also can reduce the electrical conductivity of micro-devices.

"Particles" refer to particles of any shape, any composition, and any composite structure, which can be manipulated in microfluidic devices by corresponding physical forces. An example of a microparticle is a magnetic bead that is manipulated with magnetic force. Another example of a particle is a cell manipulated with electric field forces such as traveling wave dielectrophoretic forces. The particle size used in this method can range from about 0.01 micron to about 10 cm. More suitably, the size of the particles in this method ranges from about 0.01 microns to about several thousand microns. Examples of microparticles include, but are not limited to, plastic microparticles, polystyrene microparticles, glass beads, magnetic beads, hollow glass spheres, microparticles of composite components, non-intrinsic microstructures formed by microfabrication, and the like. Other particles include cells, organelles, large biomolecules such as DNA, RNA, and proteins, among others.

"Manipulation" refers to moving and manipulating these physical molecules/microdevices, resulting in physical molecular/microdevices on a chip (including on a single chip or on or between multiple integrated chips, on a substrate or in multiple Between substrates) for one-dimensional, two-dimensional or three-dimensional motion. Manipulation of these solid molecules can also be performed in liquid containers. "Manipulating" includes, but is not limited to, transporting, focusing, enriching, concentrating, aggregating, trapping, repelling, suspending, separating, fractionating, sequestering, moving linear or other directions of the physical molecule. In order to achieve efficient manipulation, the physical molecules to be manipulated and the physical forces exerted on them should be coordinated. For example, a magnetic entity molecule can exert a magnetic force. Similarly, charged solid molecules can exert a direct current electric field force (ie, electrophoretic force). In the case of manipulating microdevices, these physical forces for manipulation of microdevices must be coordinated. For example, tiny devices are somewhat magnetic and can be manipulated by magnetic forces. Micro-devices can be composed of one or more magnetic substances, such as ferromagnetic or ferrimagnetic substances. The ferromagnetic or ferrimagnetic material can be nickel or CoTaZr (Cobalt-Tantalum-Zirconium) alloy. Similarly, tiny devices with a net charge can be manipulated using electrostatic forces. In the case of manipulation of microdevice-conjugate-entity molecule complexes, the properties of these microdevice-conjugate-entity molecules and the physical forces used for manipulation must be coordinated. For example, solid molecules or their conjugates or microdevices have certain dielectric properties, can be dielectrically polarized, and can be manipulated using dielectrophoretic forces.

"Non-directly manipulated entity molecules" means that under the action of specific physical forces, when these entity molecules are not connected to their conjugates or microdevices, no visible movement can be observed.

"Physical force" refers to such a force used to move solid molecules or their combinations or microdevices, which has no or almost no chemical or biological interactions with solid molecules or their combinations or microdevices. The reaction does not affect or hardly affects the biological and chemical properties of solid molecules or their combinations or microdevices. The term "force" or "physical force" always refers to the "force" or "physical force" acting on these solid molecules, complexes or microdevices, and the "force" or "physical force" is generated by the action of fields , determined by the properties of the physical molecule, the conjugate, or the micro-device itself. Therefore, when a field or physical field is given, in order to generate physical forces on the solid molecules, these solid molecules must have certain properties. Certain types of fields can exert forces on a variety of solid molecules with different properties, while some types of fields may only exert force on some limited types of solid molecules. For example, a magnetic field can only generate force or magnetic field force on magnetic entity molecules or entity molecules with certain magnetic properties, but is not suitable for other types of particles, such as polystyrene beads. On the other hand, a non-uniform electric field can exert physical force on many different types of solid molecules, such as polystyrene beads, cells, and magnetic beads. In fact, it is not necessary to require the physical field to have force effects on different types of solid molecules, but the physical field must be able to act on at least one solid molecule, conjugate or microdevice of at least one solid molecule, conjugate or microdevice. The effect of generating force.

"Electric field force" is the force exerted by an electric field on a solid molecule, conjugate or micro-device.

"Magnetic force" is the force exerted by a magnetic field on a solid molecule, conjugate or microdevices.

"Sound field force (sound field radiation force)" refers to the force exerted by the sound field on solid molecules, conjugates or micro-devices.

"Optical field force (optical radiation force)" is the force exerted by the optical field on solid molecules, conjugates or microdevices.

"Mechanical force" is the force exerted by a velocity field on a solid molecule, conjugate or microdevices.

"Sample" refers to any substance including physical molecules that can be separated, analyzed, manipulated, and measured by the micro-device of the present invention or the method of the present invention. The sample may be a biological sample, such as a biological fluid or biological tissue. Examples of biological fluids are urine, blood, plasma, serum, saliva, semen, feces, sputum, cerebral fluid, spinal fluid, tears, mucus, amniotic fluid, and the like. A biological tissue is an aggregate of a group of cells that can form a biological structural unit, and the tissue can be connective tissue, peripheral tissue, muscle tissue, and nervous tissue. Examples of biological tissues include organs, tumors, lymph nodes, and arteries, among others. The sample can be a target substance prepared in vitro or a mixture of enzyme solutions.

"Liquid (fluid) sample" refers to a sample in liquid or fluid form, such as a body fluid of an organism. "Liquid samples" may also refer to those samples that do not originally exist in a liquid form, such as a gas or a solid state, but are prepared in a liquid, fluid or solution. For example, a liquid sample can be prepared by dissolving or suspending tissue of a living body in a liquid (fluid).

"Determination" refers to quantitatively or qualitatively determining the composition and/or quantity of an entity molecule, such as determining the presence or absence of a certain nucleic acid or protein in a sample, and other parameters such as ratio and composition can also be determined. Assays can be performed by direct methods or by indirect methods. Measurements can be quantitative or qualitative.

Optimal Design

The present invention uses magnetic force to arrange microdevices (microdisks) into certain geometric shapes. A microdisk is a micro-machined particle with a size between 1 and 1000 microns that contains one or more strips or rods of magnetic material on one side. These strip-like or rod-like magnetic substances all have magnetization axes. The magnetization axis is determined by the geometry of the magnetic material. Typically thin films (often less than 1 micron) made of magnetic materials have an aspect ratio of no less than 3. Usually the magnetization axis of these structures is their main axis. Figure 1 shows a schematic diagram of a microdisk with two magnetic strip (rod) structures. The direction of the magnetic field is as indicated by the arrow, and the microdisk will rotate or be positioned at a certain position so that the direction of its own magnetization axis is parallel or substantially parallel to the direction of the applied magnetic field. Here, the direction of the magnetization axis of the microdisk is consistent with the direction of the main axis, or is its own main axis. If these microdisks are not restricted to a certain spatial position by other effects, they will be arranged in chains or clusters as shown in Figure 2 (the arrows in the figure indicate the direction of the magnetic field). In Figure 3, microdevices arranged in chains are confined in microchannels. Rotate the direction of the magnetic field by 90 degrees, and the microdevices arranged in a chain will rotate and be separated from each other, as shown in Figure 4. The steps shown in Figures 2 to 4 make up the "magnetic alignment".

When a magnetic field is applied, the tiny devices arrange themselves into chains or clusters. In order to introduce the microdevices into the channels, the microdevices arranged in clusters must first be dispersed. This can be achieved by rotating the direction of the magnetic field. Guide posts (discussed in detail below in the description of microchannels) can be used to provide fulcrums for rotating clusters or chains of microdevices, helping them recombine. An appropriate set of guiding posts can guide the arrangement of the microdevices into a chain, the length of the chain should be longer than the width of a single microdevice.

Chains of microdevices can be introduced into channels by magnetic force or fluid force. The chain will move in the direction of increasing magnetic field strength. If the direction of the long axis of the chain (basically consistent with the direction of the magnetization axis of the microdevice) is consistent with the direction of movement, the resistance of the liquid on the chain is relatively small, so that the chain has a faster movement speed. For a single micro-device, if the direction of the magnetization axis is perpendicular or substantially perpendicular to the direction of motion (towards the direction of increasing magnetic field strength), the micro-device will be subjected to a relatively large magnetic field force. To sum up, when the angle between the long axis direction of the chain and the direction of increasing magnetic field strength is not greater than 90 degrees, the movement of the chain is the most efficient, and the best angle is about 45 degrees. Of course, the chain Movements along other angles are also possible. Such magnetic field gradients can be achieved by using a large permanent magnet or electromagnet, or an array of small electromagnets, in or near the channel in the plane. When the magnetic field in which the microdisk is placed rotates (the direction of the magnetic field is perpendicular to the chain), the microdisk also rotates to keep in line with the change in the magnetic field.

In the present invention, it is very important to select the appropriate size of the microdisk and channel. The overlapping degree of the microdisks arranged in a chain is related to the thickness of the microdisks and the shape of the magnetic strip or rod-like structures on the microdisks. When the microdisks in FIG. 1 are arranged in a chain, there is about 20-30% overlap. When the aspect ratio of the microdisk is 1.22 (90 microns/70 microns), when the magnetic field rotates, the microdisk will also rotate in the channel, but the relative center of gravity of the microdisk in the channel will not change. There are big changes. In contrast, the disc-shaped microdisks remained overlapping as the magnetic field rotated, or were repulsed by the magnetic field and spread out on the channel's flanks.

The optimal width of the channel is determined by two factors. The channel needs to be wide enough so that a single microdevice can rotate freely in the channel. The width of the channel in Figure 1 must be at least 114 microns greater than the diagonal width of the microdisk (90 ² +70 ² square root). The minimum width of . At the same time, the channel also needs to be sufficiently narrow so that the above-mentioned micro devices do not form a string of micro devices when the main axis is perpendicular to the main axis of the micro channel. In Fig. 1, the size of the main plane of the microdisk is 90 microns X70 microns, assuming that the overlapping degree of the two microdisks is about 30%, the length of the two microdisks arranged in a chain is 153 microns (=90+90-90X30%), this is the widest width the channel can allow. When the overlapping degrees of the two microdisks are 10% and 20%, respectively, the lengths of the two microdisks arranged in a chain are 171 microns and 162 microns, respectively. To prevent the microdiscs from standing up in the channel, the height of the channel should also be taken into consideration. If the restriction of the magnetic field on the microdisk is strong enough, the situation that the microdisk stands up can be avoided, and the height of the channel itself need not be considered. The process shown in Figures 2-4 assumes that the microdisk is constrained to move on a plane. The height of the channel should be less than the length of the short side of the microdisk. If the microchannel is capped or sealed from above, the microdisk at an angle of slightly less than 90 degrees to the bottom of the channel should be able to maintain a stable upright position. The minimum angle at which the microdisk can stand stably is related to the strength of the magnetic field, the amount of magnetic substances and their saturation magnetic susceptibility, the weight and density of the microdisk and the density of the liquid medium. This value can be calculated empirically or by modeling, but generally an angle of less than 45 degrees tends to keep the microdisk horizontal in the channel. Thus, the height of the channel in which the microdisk is located must not exceed 50 microns (=70 microns×Sin45°) as shown in FIG. 1 to prevent the microdisk from remaining in a stable upright state.

The shape of the magnetic rod (strip) structure on the microdisks can affect the arrangement of the microdisks in chains or clusters, and the degree of overlap between the microdisks. Figure 5 presents a design that integrates other types of strip (rod) structures.

Minidiscs can be encoded using various methods to distinguish them from one another. The preferred coding method is to make the code on the micro-disc while micro-processing, such as a two-dimensional barcode or embedding optically recognizable characters, as shown in Figure 6. In the figure, the left side is a 2D matrix pattern, and the right side is a 2D matrix pattern. is an optically identifiable four-character coded pattern.

Encoded microdiscs can be produced using any available technology. The typical mini-disc shown in Fig. 1 comprises 4 parts: magnetic rod-like (strip) structure 1; Coding area 2 (made by aluminum material for example); Peripheral area 3 surrounds magnetic area and coding area 2, provides for Modified surface; the arrow points to the direction of the external magnetic field. The surrounding area can be made of any simple material, such as silicon-based materials, ceramics, metals, and the like. The preferred material here is silicon dioxide. These different regions are fabricated on different thickness layers of the microdisk. The magnetic rod-shaped (strip-shaped) structure 1 and the coding area 2 are fabricated in the middle layer of the microdisk, sandwiched between the materials shown in the surrounding areas 3 . These different regions are distributed at different levels of the thickness of the micro-device. The magnetic rods and coding area are located in the middle layer, wrapped by the material of the top and bottom layers. In one example of a micro-device, such a micro-disk includes a magnetic bar composed of a soft magnetic material such as CoTaZr or NiFe, measuring 90 microns long, 70 microns wide, and 3.2 microns thick.

The magnetic rod-like (strip-like) structures made in the microdisk can be made of any magnetic material. It is preferably made of a material with low remanence and high magnetic susceptibility, cobalt-tantalum-zirconium alloy meeting the above requirements. But nickel, which has a higher remanence, can also be used in the present invention. The coding layer can be made of any non-magnetic material, such as aluminum, gold or copper. Unlike the coded beads that use fluorescein as the coding method, the magnetic rod-shaped (strip-shaped) structure produced by microprocessing shown in Figure 6 is a physical code, and the number of codes that can be produced is not limited by the material itself. nature. For the 4-character encoding method shown in Figure 6, if you only take the uppercase and lowercase letters of the English alphabet and numbers (62 types in total), the number of possible encodings exceeds 10 ⁷ types.

The micro disks of the present invention can be arranged in an array, which makes it possible to quickly read the encoded information of the micro disks, and also avoids the use of complex optical detection systems. Such arrays are also amenable to long-term storage. Different from the traditional method of immobilizing capture molecules on the same surface, each different capture molecule in the microdisc array is immobilized on the surface of a microdisk. In this way, a sequence analysis can be performed on a single mini-disk. For example, after an initial screening of a certain type of disc in an array of microdiscs, the microdiscs can be further reacted with other detection molecules or subjected to another form of analysis, such as sequencing or mass spectrometry. Isolating specific microdiscs is equivalent to purifying the molecules captured on the microdiscs. In this way, when microdiscs are combined with sorting techniques, they can be used to purify solid molecules, such as proteins, DNA, cells, etc.

After entering the microchannel, the microdisks can move individually along the channel, or move as a whole in a chain. The microdiscs can be transported to the respective collection chambers through channels connected to the different collection chambers using magnetic force. For example, during DNA synthesis, each channel can be connected to one of the four reaction tubes A, T, C, and G. Such channels also allow the isolation of desired species from microdisks for subsequent analysis (see US Patent Application No. 09/924,428, filed August 7, 2001). Other sorting methods given in the Summary of the Invention herein can also be used for sorting microdiscs.

In addition to choosing a regular rectangle, the shape of the "magnetic rod-like structure" can also be a rod or other slightly irregular shape with a magnetization axis, such as an elongated cone. The examples given above are mostly planar particles (microdisks), but microdevices can take any shape including spherical beads. The simplest micro-device is an encoder with a magnetic rod-like structure. This code is made when the micro-device is made, such as by photochemical lithography or connected after the magnetic rod-like structure is made, such as by connecting a fluorescent white.

The microdevices can be arranged in an orderly form to facilitate reading the optically identifiable codes on the microdevices. The preferred arrangement of the micro-devices is to introduce the micro-devices into the channel so that the magnetization axis of the micro-devices is perpendicular to the long axis of the micro-channel, as shown in Figure 9, it is convenient to read the micro-disks arranged in chains on the glass surface The optically identifiable code of the film. As shown in Figure 10, the 2D rod-shaped code on the microdisk is fully displayed on the outside of the chain, and the light source is illuminated from below at a magnification of approximately 400 times. Although the preferred way is to have the microdisks arranged in channels (as shown in Figures 4 and 11), such an arrangement can also be done on a flat surface, such as on the surface of a glass slide (as shown in Figures 9, 10) . Even if adjacent microdisks overlap to some extent, the microdisks can be effectively arranged in a chain. "Auxiliary" microdiscs, which do not include the encoding pattern and are themselves transparent, can be added to the microdisc mix. Adding "auxiliary" microdisks before the microdisks are arranged in a chain reduces the chance of two microdisks with encoded patterns coming close. Thus, by using a magnetic field, an array of coded minidisks can be efficiently obtained.

Encoding patterns and other information are read as the particles pass through the channel. Encoding patterns and other information can be performed using any suitable sorting instrument, such as a flow cytometer.

In addition to being aligned, microdevices with magnetization axes can also rotate in a defined manner within the channel in response to changes in the external magnetic field. This rotation aids in mixing, speeding up the reaction and making the solution more homogeneous.

Example

Embodiment 1, protein profile

The surface of the coded microdisc is made of silica material, which can be silanized to bring activated functional groups on the surface, for example, use 3-aminoproplytrimethoxysilane to treat the surface to bring amino groups on the surface. Functional groups can be further activated for coupling, for example, N-hydroxysulfosuccinimide and 1-ethyl-3-(3-dimethylaminopropyl)carb-odiimide can be used to activate the amino group, and then covalently couple with the amino group on the capture antibody joint reaction. A number of these capture antibody-linked microdiscs are prepared in advance, each of which has a different biotinylated capture antibody linked to it. The mixture of such antibody-laden microdiscs is reacted with a solution containing the antigen or protein recognized by the capture antibody to be detected. After the basic reaction, add fluorescently labeled streptavidin molecules for further reaction. The microdiscs are then arrayed and the type of microdisc is detected by an optical detector while the level of antigen (protein) bound to the microdisc is determined using a fluorescence detector.

Embodiment 2, mRNA/cDNA expression profile

The surface of the encoded microdisc is made of silica material, which can be silanized to make the surface with the preferred group for attaching synthetic oligonucleotides - aldehyde groups (see "Comparison between different strategies of covalent attachment of DNA to glass surfaces to build DNA microarrays" by Zammatteo et. al. Anal. Biochem., 280: 143-150 (2000)). Amino-labeled synthetic oligonucleotides can be covalently attached to oligonucleotides by using 3-glycidoxyproplytrimethoxysilane, hydrolyzing the epoxy group to form a diol, and then treating the diol with periodate to convert the diol to an aldehyde group. The surface of the microdisk. Many such microdiscs with capture probes attached are prepared in advance, and each type of microdisk is attached with a different sequence of capture probes. The mixture of such microdiscs with capture probes is reacted with a solution containing fluorescently labeled cDNA complementary to the capture probes to be detected. After the basic reaction is complete and washed, the microdiscs are then arrayed and the type of microdisc is detected by an optical detector, while a fluorescence detector is used to determine the level of bound cDNA on the microdisc.

Embodiment 3, compound library synthesis

When there is no facility or instrument for microdisc sorting, library synthesis is random. Compound libraries can be synthesized directly on Microdiscs using the "split and pool" approach. After each step of synthesis is completed and before the next step of reaction, the microdisks are arranged for code identification. For example, when synthesizing DNA, the microdiscs are mixed, divided into four parts, and put into four reaction chambers A, G, T and C respectively. These micro-disks participating in the reaction are arranged for optical detection and record the corresponding information. Such identification and recording operations are carried out during each step of the reaction, so that the sequence of the synthesized DNA on each microdisk is recorded and known when the reaction is completed. In this method of randomly synthesizing compound libraries, the optically identifiable codes carried on each microdisk should be different, otherwise two microdisks with the same code may enter different reaction chambers to undergo different reactions, thus Different compounds are synthesized, and there is no difference in method. That is to say, in such an example, the codes on any two microdisks should be different, and two microdisks with different codes may undergo the same reaction and carry the same compound. Such a compound library can be used for screening, and such a technique can also be used for synthesizing a polypeptide library in addition to the above example for synthesizing a DNA library. Any library of compounds that can be synthesized on beads can be synthesized on such minidiscs. In combinatorial chemistry, there are a large number of such libraries (see "Comprehensive survey of combinatorial library synthesis; 1999" by Dolle Journal of Combinatorial Chemistry, 2:383-433 (2000)). This method of randomly synthesizing compound libraries requires each microdisc to have a unique code.

The second and more useful compound library synthesis requires the use of a sorting device after each step of synthesis. In this method, the types of compounds to be synthesized after synthesis are pre-specified on each microdisk before synthesis. After each reaction step, the microdisks are sorted and sent to the corresponding reaction chamber for subsequent reactions. Take a DNA molecule whose synthesis sequence is designated as ATCAGTCATGCG (SEQ ID NO: 1) for a microdisk as an example, then during the synthesis process, the microdisk first enters the test tube numbered A for synthesis, and then enters the test tube numbered T Carry out the second step of synthesis, then enter the test tube numbered C for the third step of synthesis, and so on. The library size is determined prior to synthesis, and the library to be synthesized may be only a fraction of the possible synthesized libraries. For example, a DNA library with ¹⁰⁷ kinds of 50 nucleotides in length can be synthesized, which corresponds to the sequence in the human genome, although this library is only a complete DNA ^composed of 50 nucleotides A small part of the library (10 ³⁰ species, 4 ⁵⁰ ); it is possible to synthesize a polypeptide library with 10 ⁷ kinds of 20 amino acids in length, and these 10 ⁷ kinds of polypeptide molecules correspond to the polypeptide sequences in the human genome, although this library It is only a small part of the complete 20-nucleotide polypeptide library (10 ²⁶ species, 20 ²⁰ ). Since sorting is performed at each step of the synthesis, the same compound can be produced in a single synthesis because the Microdisks with the same code will be sorted together to synthesize the same compound. This means that the number of copies on a single minidisc can be tightly controlled. What's more, the synthesized compound library can be divided into smaller parts and mixed with other libraries of known sequences to generate new libraries.

The current research hotspot in the synthesis of compound libraries is the synthesis of libraries with variable regions on templates or backbones. Many researchers and companies (such as Affibody, Phylos, Ribozyme Pharmaceuticals, Somalogic) are using this method to synthesize antibodies, enzymes, or other molecules that can recognize other molecules (such as nucleic acid antibodies, aptamers) and perform enzyme functions (such as ribozymes) or molecules that can generate signals (such as fluorescence, fluorescence energy conversion). What these methods have in common is the need to use enzymes to generate secondary libraries and interpret the final results. For example, in the screening of nucleic acid antibodies, nucleic acid antibodies are usually produced by the SELEX (Systematic Evolution of Ligands by Exponential enrichments--e.g. U.S. Pat. No. 6,048,698) process, which requires random synthesis first, and then through screening to obtain the desired A sublibrary of binding properties. This sub-library is re-amplified and randomized by PCR, and then screened. Such steps are repeated until nucleic acid antibodies that meet the pre-required specificity and affinity are obtained.

There are two major advantages to using the microdiscs of the present invention for screening and amplification. First, the amplification of the polymer does not require the use of enzymes. Second, because the polymers in the library are obtained through repeated chemical synthesis, the compounds can be any combination of the following components, nucleic acids, amino acids, small organic molecules, sugar groups, protein-nucleic acid complexes, etc. As the size of compound libraries synthesized using microdisks increases, the possibility of synthesizing molecules with special properties under extreme conditions increases, such as molecules that can capture proteins under denaturing conditions. These compound libraries can also be synthesized using traditional methods in bead format, but further screening and production are the rate-limiting steps. In traditional chemical synthesis methods, analytical chemistry methods can be used to analyze some molecules in the library, such as mass spectrometry. But it is unrealistic to analyze the properties of all the compounds in the library. But each microdisc of the present invention is optically encoded, and all components in the microdisc library can be analyzed. For example, for a library containing 10 ¹⁰ compounds, the binding efficiencies of all compounds can be analyzed, and then a new compound library can be synthesized again using a sub-library of the library as a starting point. Since the measurement information of each compound in the library is recorded in each screening process, it is convenient to use computational methods to design the next screening. When using microdisc technology, the large amount of information obtained in each round of screening of the library turns a traditional random approach into a directed systematic approach.

Claims (113)

1. a microdevice comprises: a) magnetizable base material; B) but be produced on the coding pattern of the light identification on the above-mentioned base material; C) magnetized axis.

2. according to the microdevice described in the claim 1, it is characterized in that: described magnetizable base material is paramagnetism, ferromagnetism or ferrimagnetism material.

3. according to the microdevice described in the claim 1, it is characterized in that: the absolute value of inducing the magnetization that described microdevice produces along described magnetized axis direction is induced the magnetization big at least 20% than what at least one the direction at other produced.

4. according to the microdevice described in the claim 1, it is characterized in that: the absolute value of inducing the magnetization that described microdevice produces along described magnetized axis direction is induced the magnetization big at least 50% than what at least one the direction at other produced.

5. according to the microdevice described in the claim 4, it is characterized in that: the absolute value of inducing the magnetization that described microdevice produces along described magnetized axis direction is induced the magnetization big at least 70% than what at least one the direction at other produced.

6. according to the microdevice described in the claim 5, it is characterized in that: the absolute value of inducing the magnetization that described microdevice produces along described magnetized axis direction is induced the magnetization big at least 90% than what at least one the direction at other produced.

7. according to the microdevice described in the claim 1, it is characterized in that: the absolute value of inducing the magnetization that described microdevice produces along described magnetized axis direction induces the magnetization big at least 1 times than what at least one the direction at other produced.

8. according to the microdevice described in the claim 7, it is characterized in that: the absolute value of inducing the magnetization that described microdevice produces along described magnetized axis direction induces the magnetization big at least 2 times than what at least one the direction at other produced.

9. the microdevice described in according to Claim 8 is characterized in that: the absolute value of inducing the magnetization that described microdevice produces along described magnetized axis direction induces the magnetization big at least 5 times than what at least one the direction at other produced.

10. according to the microdevice described in the claim 9, it is characterized in that: the absolute value of inducing the magnetization that described microdevice produces along described magnetized axis direction induces the magnetization big at least 10 times than what at least one the direction at other produced.

11. the microdevice according to described in the claim 10 is characterized in that: the absolute value of inducing the magnetization that described microdevice produces along described magnetized axis direction induces the magnetization big at least 20 times than what at least one the direction at other produced.

12. the microdevice according to described in the claim 11 is characterized in that: the absolute value of inducing the magnetization that described microdevice produces along described magnetized axis direction induces the magnetization big at least 50 times than what at least one the direction at other produced.

13. the microdevice according to described in the claim 1 is characterized in that: described magnetized axis is along the direction of main shaft.

14. the microdevice according to described in the claim 1 is characterized in that: described microdevice is thin discoid shape, and described magnetized axis is positioned at the principal plane of disc.

15. the microdevice according to described in the claim 1 is characterized in that: described microdevice is the thin rectangular shape shape, and the direction of described magnetized axis is along the direction of the major axis of microdevice.

16. microdevice according to claim 1 is characterized in that: the described base material that is magnetized comprises metal ingredient.

17. microdevice according to claim 16 is characterized in that: described metal ingredient is metal ingredient or its alloy material of transition group.

18. microdevice according to claim 17 is characterized in that: the metal ingredient of described transition group or its alloy material are iron, nickel, copper, cobalt, tungsten, tantalum, zirconium or nickel-ferro alloy and cobalt-tantalum-zircaloy.

19. microdevice according to claim 16 is characterized in that: described metal ingredient is Fe3O4.

20. microdevice according to claim 1 is characterized in that: described microdevice comprises the substrate that a non-magnetisable material is made.

21. the microdevice according to described in the claim 20 is characterized in that: described base material is silica-base material, plastics, pottery, rubber, polymer or their compound.

22. the microdevice according to described in the claim 21 is characterized in that: described silica-base material is silicon dioxide or silicon nitride.

23. the microdevice according to described in the claim 20 is characterized in that: the surface of described substrate is hydrophobic or hydrophilic.

24. the microdevice according to described in the claim 1 is characterized in that: but the composition of described light recognition coding pattern by the material of described microdevice own provides, the structure of described microdevice itself provides or use the optical markings thing to provide.

25. the microdevice according to described in the claim 24 is characterized in that: but the diversity of described light recognition coding pattern is made up of shape, quantity, position distribution, chemical property and the material of the structure of substrate, substrate and optical markings thing or their combining case provides.

26. the microdevice according to described in the claim 24 is characterized in that: but the structure that described light recognition coding pattern is one group of microdevice is arranged or one group of optical markings thing.

27. the microdevice according to described in the claim 1 is characterized in that: but described light recognition coding pattern is to use general processing technology or micro fabrication is produced on the microdevice.

28. the microdevice according to described in the claim 1 is characterized in that: but described light recognition coding pattern is to use the mask etching fabrication techniques.

29. the microdevice according to described in the claim 28 is characterized in that: described mask etching technology is photoengraving, electron beam lithography or X ray etching.

30. the microdevice according to described in the claim 24 is characterized in that: described optical markings thing is placed on the described microdevice.

31. the microdevice according to described in the claim 24 is characterized in that: described optical markings thing is the part of described microdevice.

32. the microdevice according to described in the claim 24 is characterized in that: described optical markings thing is vapour deposition or sputters on the microdevice.

33. the microdevice according to described in the claim 24 is characterized in that: but described optical markings thing is particulate or quantum dot that the fluorescent material scattered light detects.

34. the microdevice according to described in the claim 33 is characterized in that: described quantum dot is cadmium-X (Cd-X) core texture, and wherein X represents selenium (Se), sulphur (S) or tellurium (Te).

35. the microdevice according to described in the claim 34 is characterized in that: described quantum dot is realized passivation by parcel one deck inorganics.

36. the microdevice according to described in the claim 35 is characterized in that: described integument is a Y-Z type structure, and wherein Y represents cadmium or zinc, and Z represents sulphur or selenium.

37. the microdevice according to described in the claim 33 is characterized in that: described quantum dot comprises a cadmium-X (Cd-X) core texture, and wherein X represents selenium (Se), sulphur (S) or tellurium (Te); With a Y-Z type integument, wherein Y represents cadmium or zinc, and Z represents sulphur or selenium; In addition, the skin of described microdevice wraps up one deck trialkyl phosphine oxide again.

38. the microdevice according to described in the claim 24 is characterized in that: but described light recognition coding pattern is the bar coding of one dimension and/or two dimension.

39. the microdevice according to described in the claim 1 is characterized in that: described microdevice also comprises the bond that can combine with entity molecule to be handled.

40. the microdevice according to described in the claim 39 is characterized in that: described bond is antibody or nucleotide sequence.

41. the microdevice according to described in the claim 39 is characterized in that: described microdevice has one group of bond, and every kind of bond can be special in conjunction with a kind of entity molecule to be handled.

42. the microdevice according to described in the claim 1 is characterized in that: described microdevice has one and is convenient to microdevice/entity molecule-microdevice compound manipulation or feasible to microdevice/entity molecule-possible unit of microdevice compound manipulation becoming.

43. the microdevice according to described in the claim 42 is characterized in that: described unit is conductive material, insulating material, have the material and the charged materials of high/low acoustic damping character.

44. the microdevice according to described in the claim 42 is characterized in that: described unit is handled by dielectrophoresis force, row ripple dielectrophoresis force, sound field power, electrostatic force, mechanical force, optical radiation power or thermal convection power.

45. the microdevice according to described in the claim 15 is characterized in that: the main shaft of described microdevice and the ratio of secondary axes are at least 1.2.

46. the microdevice according to described in the claim 15 is characterized in that: described microdevice comprises the rectangular configuration that two paramagnets are made at least.

47. the microdevice according to described in the claim 46 is characterized in that: described rectangular configuration is separated by metal level.

48. the microdevice according to described in the claim 47 is characterized in that: described metal level comprises aluminium.

49. the microdevice according to described in the claim 47 is characterized in that: described microdevice has the rectangular configuration that the different paramagnet of quantity is made in the main shaft both sides.

50. the microdevice according to described in the claim 15 is characterized in that: described microdevice has the rectangular configuration that two paramagnets are made along main shaft.

51. the microdevice according to described in the claim 46 is characterized in that: described paramagnet forms the rectangular configuration along the major axes orientation of described microdevice, and this rectangular configuration all has the outstanding structure of finger-shaped at two ends.

52. the microdevice according to described in the claim 50 is characterized in that: the rectangular configuration that described two paramagnets are made all has the outstanding structure of finger-shaped at two ends.

53. the microdevice according to described in the claim 1 is characterized in that: described microdevice also comprises in order to synthetic, combination and the functional group that is connected usefulness.

54. the microdevice according to described in the claim 53 is characterized in that: described functional group is carboxyl, amino, hydroxyl, sulfydryl, epoxy radicals, ester group, thiazolinyl, alkynyl, alkyl, aromatic radical, aldehyde radical, ketone group or their derivant.

55. the microdevice according to described in the claim 24 is characterized in that: the structure of described microdevice itself is the hole of getting on described microdevice.

56. the microdevice according to described in the claim 46 is characterized in that: the rectangular configuration that described at least two paramagnets are made is arranged in strip.

57. the system that can constitute the microdevice array comprises a) microdevice described in one group of claim 1; B) the minitype channel array of forming by one group of minitype channel; The width of described minitype channel is: when described microdevice was in the externally-applied magnetic field, single microdevice can rotate freely in passage, can not form the microdevice chain when the main shaft of described microdevice main shaft and minitype channel is vertical.

58. the system according to described in the claim 57 is characterized in that: the height of described minitype channel and/or the restriction of microdevice is made that microdevice is unlikely to erect in minitype channel by magnetic field.

59. the system according to described in the claim 57 is characterized in that: the height of described minitype channel is not more than 70% of microdevice main axis length.

60. the system according to described in the claim 57 is characterized in that: described minitype channel array comprises one can be so that the zone that microdevice is handled therein.

61. the system according to described in the claim 57 is characterized in that: described minitype channel array comprises one in order to discharge the exit passageway of microdevice.

62. the system according to described in the claim 57 is characterized in that: described minitype channel array comprises the field generator for magnetic that can handle microdevice turnover minitype channel.

63. the system according to described in the claim 62 is characterized in that: described field generator for magnetic is ferromagnetic material or little electromagnetic unit.

64. the system according to described in the claim 62 is characterized in that: described field generator for magnetic below the described minitype channel array, middle, above and near.

65. a method of making the microdevice array may further comprise the steps:

(a) provide one group of microdevice described in the claim 1;

(b) provide a minitype channel array of forming by one group of minitype channel; The width of described minitype channel is: when described microdevice was in the externally-applied magnetic field, single microdevice can rotate freely in passage, can not form the microdevice chain when the main shaft of described microdevice main shaft and minitype channel is vertical;

(c) described one group of microdevice is introduced described minitype channel;

(d) rotate described microdevice by the magnetic field force that adds in minitype channel, the acting in conjunction of the magnetized axis by described magnetic field force and described microdevice makes and disperses mutually between the described microdevice.

66. the method according to described in the claim 65 is characterized in that: the height of described minitype channel and/or the restriction of microdevice is made that microdevice is unlikely to erect in minitype channel by magnetic field.

67. the method according to described in the claim 65 is characterized in that: the height of described minitype channel is not more than 70% of microdevice main axis length.

68. the method according to described in the claim 65 is characterized in that: described microdevice is introduced minitype channel by magnetic field force, fluid force or both combinations.

69. the method according to described in the claim 65 is characterized in that: described microdevice is introduced minitype channel by magnetic field force, and the direction of introducing makes the angle between microdevice main shaft and the minitype channel main shaft be not more than 45 degree.

70. the method according to described in the claim 65 is characterized in that: described method is included in minitype channel the microdevice of introducing before the microdevice or interrupting catenation when introducing.

71. the method according to described in the claim 65 is characterized in that: the rotation of described microdevice is no less than 45 degree.

72. the method according to described in the claim 71 is characterized in that: described microdevice revolves and turn 90 degrees.

73. the method according to described in the claim 65 is characterized in that: at least one is in conjunction with a kind of steerable entity molecule in described one group of microdevice.

74. the method according to described in the claim 65 is characterized in that: described one group of microdevice is respectively in conjunction with one group of steerable entity molecule.

75. the method according to described in the claim 73 is characterized in that: described manipulation be transport, focusing, enrichment, concentrate, assemble, catch, the moving of the entity molecule on repulsion, suspension, separation, fractionation, isolation or the linear direction.

76. the method according to described in the claim 73 is characterized in that: but thereby described method comprises the coding pattern that uses optical means to detect light identification on the microdevice determines the entity molecule of being handled on this microdevice.

77. the method according to described in the claim 65 is characterized in that: described one group of microdevice combines with a group object molecule; Described method realizes the detection by quantitative to entity molecule by the physical parameter of the described entity molecule of detection or by detecting and the interaction of described entity molecule or the physical parameter of the mark-up entity molecule that is connected.

78. the method according to described in the claim 77 is characterized in that: the physical parameter of described physical parameter or mark-up entity molecule is that fluorescence, radioactive isotope, quality, reflectivity, absorptivity, chemiluminescence or described parameter pair interact or the response of the mark-up entity molecule that is connected with described entity molecule.

79. the method according to described in the claim 78 is characterized in that: described and described entity molecule interacts or the mark-up entity molecule that is connected is an enzyme.

80. the method according to described in the claim 73 is characterized in that: described method also comprises by collect the step of the microdevice that combines entity molecule at endpiece.

81. the method described in 0 according to Claim 8 is characterized in that: described method also comprises the step that discharges entity molecule from the microdevice of collecting.

82. a method of making the microdevice array may further comprise the steps:

(a) provide one group of microdevice described in the claim 1, they are positioned on the plane that is beneficial to microdevice rotation;

(b) rotate described microdevice by the magnetic field force that adds on described surface, the acting in conjunction of the magnetized axis by described magnetic field force and described microdevice makes and disperses mutually between the described microdevice.

83. the method described in 2 is characterized in that: be manufactured with width on the described surface and be not more than the described microdevice groove structure of the width of one dimension at least according to Claim 8.

84. the method described in 2 according to Claim 8, it is characterized in that: described method comprises introduces described surface with described microdevice with the form of liquid suspension, removes the step of described liquid then by groove.

85. the method in a synthetic compound storehouse may further comprise the steps:

1) provide one group of microdevice, this microdevice comprises: (a) base material that can be magnetized; (b) but be produced on the coding pattern of the light identification on the above-mentioned base material; (c) magnetized axis; But the coding pattern correspondence of described light identification the product that will carry out synthetic reaction on described microdevice;

2) synthetic described compound library: synthetic reaction realizes by a series of synthesis steps; In per step synthetic in, but according to the light recognition coding pattern on the described microdevice, carry out corresponding synthetic reaction; Behind the single step reaction, but, described microdevice is classified, carry out next step synthetic reaction more again according to the light recognition coding pattern on the described microdevice.

86. the method described in 5 is characterized in that: the minitype channel array sorting of described microdevice by being made of one group of minitype channel according to Claim 8; The width of described minitype channel is: when described microdevice was in the externally-applied magnetic field, single microdevice can rotate freely in passage, can not form the microdevice chain when the main shaft of described microdevice main shaft and minitype channel is vertical; The acting in conjunction of the magnetized axis by the magnetic field force that adds and described microdevice makes between the described microdevice and disperses mutually.

87. the method described in 5 according to Claim 8 is characterized in that: the height of described minitype channel and/or the restriction of microdevice is made that described microdevice is unlikely to erect in described minitype channel by magnetic field.

88. the method described in 5 according to Claim 8, it is characterized in that: the height of described minitype channel is not more than 70% of microdevice main axis length.

89. the method described in 5 is characterized in that: connected a kind of synthetic compound on described each microdevice according to Claim 8.

90. the method described in 5 according to Claim 8, it is characterized in that: described synthetic compound is peptide, protein, nucleic acid oligomer, nucleic acid, vitamin, oligosaccharides, carbohydrates, fat, micromolecule or their compound.

91. 5 described methods according to Claim 8, it is characterized in that: described compound library comprises one group of entity molecule relevant with a certain biological pathway, this group object molecule has identical or similar biological function, all express in the same cell cycle, in same cell type, express, in same types of organization, express, in same organ, express, in the same stage of development, express; The expression of entity molecule can be owing to disease changes or active, or changes along with disease type and advancing of disease stage, or is subjected to medicine or other processing and changes.

92. 5 described methods according to Claim 8, it is characterized in that: described synthetic compound library comprises one group of nucleic acid fragment.

93. according to the described method of claim 92, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 2 nucleotide at least.

94. according to the described method of claim 93, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 10 nucleotide at least.

95. according to the described method of claim 94, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 15 nucleotide at least.

96. according to the described method of claim 95, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 20 nucleotide at least.

97. according to the described method of claim 96, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 25 nucleotide at least.

98. according to the described method of claim 97, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 50 nucleotide at least.

99. according to the described method of claim 98, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 75 nucleotide at least.

100. according to the described method of claim 99, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 100 nucleotide at least.

101. according to the described method of claim 100, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 200 nucleotide at least.

102. according to the described method of claim 101, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 500 nucleotide at least.

103. 5 described methods according to Claim 8, it is characterized in that: described synthetic compound library comprises a histone matter or a polypeptide fragments.

104. according to the described method of claim 103, it is characterized in that: protein or polypeptide fragments in the described synthetic compound library comprise 2 amino acid at least.

105. according to the described method of claim 104, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 10 amino acid at least.

106. according to the described method of claim 105, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 15 amino acid at least.

107. according to the described method of claim 106, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 20 amino acid at least.

108. according to the described method of claim 107, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 25 amino acid at least.

109. according to the described method of claim 108, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 50 amino acid at least.

110. according to the described method of claim 109, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 75 amino acid at least.

111. according to the described method of claim 110, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 100 amino acid at least.

112. according to the described method of claim 111, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 200 amino acid at least.

113. according to the described method of claim 112, it is characterized in that: the nucleic acid fragment in the described synthetic compound library comprises 500 amino acid at least.

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