TWI611185B - Detection device - Google Patents

Abstract

A detection device includes a substrate, a cover, a micro-sensing wafer, and a micro-channel structure, wherein at least one of the substrate and the cover includes a porous material. The substrate has a first surface with a recessed portion on the first surface. The recessed portion includes a bottom and a slope. The bottom is embedded in the substrate. The slope connects the first surface and the bottom and is disposed at one end of the recessed portion near an injection port. The cover has a second surface facing the first surface. The micro-sensing wafer is embedded in the substrate. The microchannel structure is embedded in the second surface and the first surface to form a microchannel. A specimen enters the depression from the injection port through the microchannel and is separated into a lower layer liquid staying at the bottom and an upper layer liquid flowing from the depression to The micro-sensing wafer is used, and no extra power is applied to the detection device during the detection of the specimen.

Description

Detection device

The present invention relates to a detection device, in particular to a detection method that separates a heavier molecule from a lighter molecule in a specimen and carries out a related inspection at the same time without the need for additional power during the inspection process. Device.

In the existing microfluidic detection device, a capillary phenomenon formed by the difference between the adhesion between the specimen and the microfluidic channel and the cohesive force of the specimen itself enables the specimen to flow in the microfluidic channel. After the specimen passes a test area in the microfluidic channel, the micro-sensing wafer in the test area can detect the specimen. Finally, the researchers transmit the detection signals read by the micro-sensing chip to an external analysis instrument for related research and data analysis. However, current detection devices must be powered by microfluids in the microfluidic channels, which greatly reduces the portability of the detection devices.

In some specific specimen detection procedures, researchers need to screen the specimen to obtain a specific molecule in the specimen, and then analyze and study the specific molecule. For example, when the specimen is blood, in order to separate blood cells from plasma, the conventional detection method uses the characteristics of different qualities of blood cells and plasma, and separates the blood into blood cells and plasma with a centrifugal separator. Therefore, adopting the above-mentioned conventional techniques not only has complicated procedures and lengthy testing time, but also requires the power of a centrifugal separator for testing, which is very inconvenient.

In view of this, the object of the present invention is to provide a detection device that can directly separate a sample into a molecule with a heavier mass and a molecule with a lighter mass, and perform detection on a sample with the separated molecule, so there is no need to perform centrifugal separation. Separating the sample from the machine not only simplifies the testing procedure, but also has a green concept of energy saving.

Another object of the present invention is to provide a detection device which is highly portable and does not require external power during the detection of a specimen.

In order to achieve the above object, the detection device of the present invention includes: a substrate having a first surface, the first surface having a recessed portion, the recessed portion includes a bottom and a slope, the bottom is embedded in the substrate, and the slope connects the first surface with The bottom is disposed at one end of the recess; a cover having a second surface facing the first surface; a micro-sensing wafer embedded in the substrate; and a micro-channel structure embedded in the second surface, wherein the cover covers After the substrate, the first surface and the second surface are closely adhered to each other, so that the microfluidic channel structure and the first surface are connected to form a microfluidic channel including at least one injection port and a capacity control groove to control the flow of a specimen in the microfluidic channel. The sample enters the depression from the injection port through the microchannel, and the sample is separated into a lower layer liquid and an upper layer liquid at the depression portion, the lower layer liquid stays at the bottom, and the upper layer liquid flows from the depression portion to the micro-sensing chip; The micro-channel includes a first-class choke channel and a reaction tank. The reaction tank is connected to the micro-sensing chip, and a choke channel is formed between the reaction tank and the capacity control tank. Wherein the lid comprises one of a porous material.

In one embodiment of the present invention, the specimen is blood, the lower fluid is blood cells, and the upper fluid is plasma.

In an embodiment of the present invention, one end of the micro-flow channel has a capacity control slot, which can control the flow rate of the specimen in the micro-flow channel.

In an embodiment of the present invention, the above-mentioned slope is disposed in the recessed portion near one end of the injection port.

In one embodiment of the present invention, the micro-sensing wafer has at least one detection structure, and the detection structure energizes biological particles or biopolymers in the specimen.

In an embodiment of the present invention, the above-mentioned detection device further includes a plurality of terminals, which are disposed on the substrate and connected to the micro-sensing chip, and the plurality of terminals can be coupled to a reading device.

In an embodiment of the present invention, the plurality of terminals are connected to the micro-sensing chip in a wire bonding manner.

In an embodiment of the present invention, the aforementioned detection structure is a resistance type, a capacitance type, an impedance type, or a transistor type, or an electrochemical type, or a counting type, or a photoelectric type using a nanometer sensing material as a basis. In the sensor, the nanomaterial is functionalized by a biopolymer, which is selected from antibodies, aptamers or sugar molecules or enzymes. In addition to the nano-sensing material as the basis, the detection structure can also choose purely electrochemical or photoelectric sensors.

In one embodiment of the present invention, the nanometer sensing material is selected from carbon nanotubes, graphene, reduced graphene oxide (rGO), and graphene oxide (graphene). oxide, GO), nanoribbon graphene, nano silicon wire, nano InP wire, nano GaN wire, nano semiconductor wire or nano semiconductor film.

In an embodiment of the present invention, the material of the substrate is polymethylmethacrylate (PMMA), polyethylene terephthalate (PET), polycarbonate (PC), and porosity. Polydimethylsilicon (PDMS), porous silicone, rubber, plastic or glass.

In an embodiment of the present invention, the material of the cover body is polymethylmethacrylate (PMMA), porous polyethylene terephthalate (PET), and polycarbonate (PC). ), Porous polydimethylsilicon (PDMS), porous silicone, rubber or plastic.

In one embodiment of the present invention, a pre-processing portion is provided between the recessed portion and the injection port, which is suitable for separating the sample or mixing it with other reagents.

In an embodiment of the present invention, when the second surface is assembled with the first surface, an edge of the second surface is located inside the edge of the first surface.

In an embodiment of the present invention, the detection device after the completion of the evacuation procedure is packaged in a vacuum packaging bag.

In an embodiment of the present invention, the first surface and the top surface of the micro-sensing chip are in the same plane.

In an embodiment of the present invention, the above-mentioned microchannel is located in a region between the micro-sensing wafer and the recessed portion, and the cross-sectional area of the channel in the microchannel is reduced.

To achieve the above object, a detection device of the present invention includes: a substrate having a first surface including a recessed portion and at least one reaction tank; a cover body having a second surface covering the first surface of the cover body; The micro-sensing wafer is embedded in the substrate and includes at least one sensing area; a first injection port; a second injection port; and a micro-channel system formed between the substrate and the cover, including a first micro-channel formed on the substrate. The first surface communicates with the reaction tank and the second injection port, a second microchannel is formed on the second surface of the cover, and a capacity control channel is formed on the second surface of the cover and communicates with the second microchannel And a third microfluidic channel connecting some reaction tanks to the micro-sensing chip At least one sensing area; wherein the second microfluidic channel includes a first part and a second part, the first part extends from the first injection port to the capacity control slot and communicates with the recessed part, and the second part is formed by The capacity control tank extends to at least one reaction tank; wherein at least one of the substrate and the cover body comprises a porous material.

To achieve the above object, a detection device of the present invention includes: a substrate having a first surface including a recessed portion and a decomposition groove; a cover body having a second surface covering the first surface of the cover body; a micro The sensing chip is embedded in the substrate and includes a sensing area; a microfluidic channel is formed between the substrate and the cover, and communicates with the recessed portion and the sensing area of the microsensing chip; a heating component is formed below a part of the microfluidic channel; Among them, at least one of the substrate and the cover body includes a porous material; one of the decomposition solutions contains a DNA-containing sample, which is filled with the decomposition tank and the depression, and then flows through the microchannel to the heating component, and is heated by the heating component in a large amount. Duplicate, and then flow to the sensing area of the micro-sensing chip through the micro-channel.

In order to make the above and other objects, features, and advantages of the present invention more comprehensible, embodiments are described in detail below with reference to the accompanying drawings.

10, 20, 30‧‧‧ detection devices

100, 700‧‧‧ substrate

101‧‧‧first surface

102, 702‧‧‧ Depression

103‧‧‧ bottom

104‧‧‧ slope

200, 800‧‧‧ cover

300, 900‧‧‧ micro-sensing chip

814‧‧‧first sensing cavity

816‧‧‧Second sensing cavity

818‧‧‧third sensing cavity

820‧‧‧Fourth sensing cavity

400‧‧‧Micro channel structure

401, 802, 804, 704‧‧‧ microchannel

402‧‧‧Injection port

403, 803‧‧‧Quantitative control structure

404‧‧‧Pretreatment Department

405, 805‧‧‧ Capacity control slot

408, 808‧‧‧ flow choke

409‧‧‧ reaction tank

500、720‧‧‧Terminal carrier board

501, 721‧‧‧ terminal

600‧‧‧ specimen (blood)

601‧‧‧ Underlayer (blood cells)

602‧‧‧ upper fluid (plasma)

603‧‧‧Biomarker

604‧‧‧ aptamer

706‧‧‧The first reaction tank

708‧‧‧Second reaction tank

710‧‧‧Third reaction tank

712‧‧‧Fourth reaction tank

750‧‧‧Heating component

751‧‧‧Heating wafer

752‧‧‧ resistance line

752a, 752b‧‧‧ conductive terminal

760‧‧‧Decomposition tank

809‧‧‧sensing cavity

810‧‧‧first injection port

812‧‧‧Second injection port

FIG. 1 is a schematic perspective view of a detection device according to an embodiment of the present invention.

FIG. 2 is a schematic perspective view of a substrate of a detection device according to an embodiment of the present invention.

FIG. 3 is a schematic perspective view of a recessed portion according to an embodiment of the present invention.

FIG. 4 is a schematic three-dimensional exploded view of a substrate and a cover of a detection device according to an embodiment of the present invention.

FIG. 5 is a schematic three-dimensional exploded view of a substrate and a cover of a detection device according to another embodiment of the present invention.

FIG. 6 is a schematic view of a specimen in a recessed portion according to an embodiment of the present invention.

FIG. 7 is a schematic diagram showing a reduction in a cross-sectional area in a microchannel according to an embodiment of the present invention;

FIG. 8 is a schematic diagram of a pre-processing section according to an embodiment of the present invention.

FIG. 9 is a test record table of a detection device according to an embodiment of the present invention.

FIG. 10 is a schematic perspective view of another embodiment of the present invention.

11A and 11B are schematic top views of another embodiment of the present invention.

12 to 16 show different embodiments of the recessed portion of the substrate according to another embodiment of the present invention.

Fig. 17 is a top perspective view showing another embodiment of the present invention.

FIG. 18 is a top perspective view showing another embodiment of the detection device of the present invention.

Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, the word "a" means the quantity of at least one (one or more) unless otherwise specified.

FIG. 1 is a perspective view of a detection device according to an embodiment of the present invention, FIG. 2 is a perspective view of a substrate of a detection device according to an embodiment of the present invention, and FIG. 3 is a depression according to an embodiment of the present invention. FIG. 4 is a three-dimensional exploded view of a substrate and a cover of a detection device according to an embodiment of the present invention, and FIG. 5 is a three-dimensional exploded view of a substrate and a cover of a detection device according to another embodiment of the present invention . Please refer to FIG. 1, FIG. 2, FIG. 3, FIG. 4, and FIG. 5. The detection device 10 of the present invention includes a substrate 100, a cover 200, a micro-sensing wafer 300, and Micro-channel structure 400. The substrate 100 has a first surface 101, and the first surface 101 has a recessed portion 102. The recessed portion 102 is composed of a bottom portion 103 and a slope 104. The bottom portion 103 is embedded in the substrate 101, and the slope 104 is located at one end of the recessed portion 102 near the injection port 402.

Further, in the embodiment of the present invention, at least one of the substrate 100 and the cover 200 includes a porous material, and the cover 200 has a second surface 201 thereon. When the cover 200 covers the substrate 100, the first surface 101 faces the second surface 201, and the two are in close contact with each other. The second surface 201 has a microchannel structure 400. When the cover 200 covers the substrate 100, the microchannel structure 400 is combined with the first surface 101 to form a microchannel 401. When at least the substrate 100 and the cover 200 are in it, After one of the porous materials forms a vacuum state with the inside of the microchannel 401, the specimen 600 is placed at the injection port 402 of the microchannel 401, and the specimen 600 is driven from the injection by the suction generated by the vacuum in the microchannel 401. The inlet 402 enters the recessed portion 102 through the microchannel 401, and the specimen 600 is separated into the lower layer liquid 601 and the upper layer liquid 602 (as shown in FIG. 6) at the recessed portion 102, and the lower layer liquid 601 stays at the bottom 103 and the upper layer liquid. 602 flows from the depression 102 to the micro-sensing wafer 300.

In the embodiment of the present invention, the specimen 600 may refer to body fluid, including blood, cerebrospinal fluid, gastric fluid, and various digestive fluids, semen, saliva, tears, sweat, urine, vaginal secretions, or the like. A solution containing the specimen 600. In this embodiment, blood is taken as an example. When blood enters the depression 102 along the microchannel 401, the heavier blood cells (lower layer liquid 601) will precipitate on the bottom 103 of the depression 102, and the lighter plasma ( The upper layer liquid 602) exits from the recess 102 and enters the micro-sensing wafer 300 along the micro-channel 401. When the specimen 600 is thick, such as blood, in order to make the specimen 600 more smoothly injected from the injection port 402, the bottom and / or the side wall of the injection port 402 may be coated with an anticoagulant.

It is worth mentioning that the slope 104 of the recessed portion 102 of the present invention has a secure specimen 600 can perform the separation function in the recessed portion 102. In the conventional technology, the specimen 600 often has a poor separation effect, which results in a misjudgment result when the separated specimen 600 is read by the micro-sensing wafer 300. In the detection device 10 of the present invention, the slope 104 of the end of the recessed portion 102 near the injection port 402 has a function of smoothing the specimen 600 when flowing in the microchannel 401. When the specimen 600 is in the microchannel 401, When flowing smoothly to the recessed portion 102, molecules of different weights can produce different sinking speeds according to different weights with relatively little external interference. Therefore, molecules of different weights can be more efficiently separated . In order to increase the effect of separating the specimen 600 from the depression 102, micro-pillars arranged in an array may be formed at the junction of the depression 102 and the microchannel 401. The spacing between the micro pillars is less than 3 microns to intercept the suspension above 3 microns. Or form micro-cylinders with a distance between 10 to 100 microns, and use a plurality of microspheres with a size larger than the distance between the plurality of micro-cylinders to form a plurality of voids to intercept suspended matter larger than the size of the void. The inclination and / or surface roughness of the slope 104 of the depression 102 can intercept more suspended matter. The surface of the bottom 103 and the slope 104 of the depression 102 may be treated with an oxygen plasma or a surfactant to increase hydrophilicity and increase the probability of the suspended matter in the specimen 600 to settle. If the specimen 600 is blood containing platelets, a coagulant such as calcium chloride (CaCl) may be coated on the bottom 103 and the slope 104 of the depression 102 to promote blood cell coagulation and aggregation and settle in the depression 102. If the specimen 600 is, for example, a non-blood specimen of somatic cells in raw milk, it may be coated on the bottom 103 and the slope 104 of the depression 102 or a mixture including thrombin (thrombin) and fibrinogen may be added to the specimen 600. (fibrinogen) and calcium ions (Calcium ion) to form a fibrin mesh, which increases the probability of the suspended matter settling in the depression 102.

When the sample 600 is in the process of separation, the research project usually includes quantitative analysis of the separated sample. In the embodiment of the present invention, the micro-channel 401 is opposite to the injection port. The other end of 402 has a capacity control slot 405, the purpose of which is to design a structure designed for quantitative sample 600 analysis. After the specimen 600 enters the microchannel 401 from the injection port 402, it passes through the recess 102 and the micro-sensing wafer 300, and finally the specimen 600 is stored in the capacity control tank 405. When the sample 600 is filled with the capacity control slot 405, the sample 600 at the injection port 402 will not enter the microchannel 401, so the signal detected by the micro-sensing chip 300 is contained in the capacity control slot 405. The signal generated by the quantitative sample 600. In this embodiment, it is assumed that the capacity control tank 405 has a capacity of 0.5 cc. Although the sample 600 applied to the injection port 402 is much larger than 0.5 cc, the signal sample 600 that can be detected by the micro-sensing chip 300 is only 0.5. cc. If the signal detected by the micro-sensing chip 300 is divided by 0.5 cc, the unit of the signal is presented as a concentration method. In this embodiment, a certain amount of control structure 403 is provided on the second surface 201, and the capacity control groove 405 can be formed by combining the first surface 101 and the quantitative control structure 403.

In an embodiment of the present invention, the micro-sensing wafer 300 is embedded in the substrate 100, and the top surface of the micro-sensing wafer 300 and the first surface 101 must be the same plane to ensure that the specimen 600 in the micro-channel 401 can be Flow into the micro-sensing wafer 300. In this embodiment, the microfluidic channel 401 passes through at least one detection structure of the micro-sensing wafer 300 from above. In another embodiment, the micro-fluidic channel 401 can also pass through at least one detection structure of the micro-sensing wafer 300 from below. . Each detection structure is modified by biological coupling, which can quantify the biological particles or biopolymers in the specimen 600, and can further pass the micro-sensing chip 300 such as a resistive type, a capacitive type, an impedance type, or a transistor type, Or the electrochemical type contains nano or non-nano, or the counting type, the photoelectric type contains nano or non-nano sensing elements, and converts them into electrical signals. Finally, the I / O pads of the micro-sensing chip 300 are electrically connected. The plurality of terminals 501 are electrically connected to the external reading device through the plurality of terminals 501, and the detection signals are output to provide relevant research and analysis. In the embodiment of the present invention, the plurality of terminals 501 may also be used for punching. The wire bonding is connected to the micro-sensing chip 300. In some embodiments, the micro-sensing chip 300 may also include an amplifier circuit to amplify the weak electronic signal detected.

When measuring the specimen 600 using the detection device 10, in order to detect the low-concentration specimen 600, the present invention designs a micro-fluid channel 401 in a region where the micro-fluid channel 401 is located between the micro-sensing wafer 300 and the recessed portion 102. The internal cross-sectional area is reduced, so that the flow rate of the specimen 600 entering the micro-sensing wafer 300 is reduced. This can increase the time that the specimen 600 stays in the micro-sensing wafer 300, and can make most of the specimens 600 closer to the micro-sensing wafer. 300 to facilitate the detection of low-concentration specimen 600. As shown in FIG. 7, in an embodiment of the present invention, the channel depth of the micro channel 401 was originally 60 μm, and the channel depth between the micro-sensing chip 300 and the recessed portion 102 can be gentle with a slope. Reduced to 10 μm, so that the biomarker 603 in the deep flow channel (60 μm) upstream of the micro flow channel 401 can therefore pass through the slope, slow its flow velocity, limit its suspension range, and therefore rush to the aptamer 604 at the bottom of the micro flow channel 401. As a result, most of the biomarkers 603 are captured by the aptamer 604 at the bottom of the microfluidic channel 401. Because of the low flow rate, any captured biomarkers 603 will be fixed on the aptamer 604.

In the detection device 10 of the present invention, the detection structure is a resistance-type, capacitance-type, impedance-type, or transistor-type, electrochemical-type, or counting-type sensor based on a nano-sensing material. The test material is functionalized with a biopolymer, which means at least one of an antibody, an aptamer, a sugar molecule, or an enzyme molecule. The sensor may be a plurality or an array type to provide quantitative inspection of a variety of targets in the specimen 600. The nano-sensing material described above may be a material having semiconductor characteristics such as a nanowire suitable for sensing, such as a carbon nanotube, a nano-silicon wire, a nano-InP wire, a nano-GaN wire, or Nanometer semiconductor wires, or nanometer semiconductor thin films, or graphene, reduced graphene oxide (rGO), graphene oxide (GO), Nanoribbon graphene and the like. In addition to the nano-sensing material as the basis, the detection structure can also choose purely electrochemical or photoelectric sensors.

In the embodiment of the detection device 10 of the present invention, the material of the substrate 100 may be acrylic (polymethylmethacrylate, PMMA), polyethylene terephthalate (PET), or polycarbonate (PC). Porous polydimethylsilicon (PDMS), porous silicone, rubber, plastic or glass; the material of the cover 200 may be acrylic (polymethylmethacrylate, PMMA), polyethylene terephthalate Polyester (polyethylene terephthalate, PET), polycarbonate (PC), porous polydimethylsilicon (PDMS), porous silicone, rubber or plastic.

It should be mentioned that when selecting materials for the substrate 100 and the cover 200, the material characteristics between the substrate 100 and the cover 200 must be considered. When the cover body 200 covers the substrate 100, the inside of the microfluidic channel 401 must be evacuated to form a vacuum state to provide suction for driving the specimen 600 to flow in the microfluidic channel 401. Therefore, at least one of the substrate 100 and the cover 200 must be a porous material. Preferably, the hardness characteristics of the material between the substrate 100 and the cover 200 can be hard and soft. Taking one embodiment of the present invention as an example, the material of the substrate 100 is made of plastic with higher hardness, and the cover 200 is made of porous polydimethylsilicon (PDMS) with lower hardness. The micro-channel structure 400 is formed on the low cover 200, so when the micro-channel 401 is evacuated, the porous polydimethylsilicon (PDMS) with lower hardness will be attached to the hardness On the high plastic substrate 100, the air in the pores of the PDMS will be evacuated to maintain the vacuum state in the microchannel 401. Pore vacuum state of porous polydimethylsilicon (PDMS) when the detection device 10 is exposed to normal atmospheric pressure It will gradually be filled with outside air, but as long as it is not in equilibrium with the outside pressure, a negative pressure of the microchannel 401 can be provided to drive the specimen 600 to flow.

In the above embodiment, the substrate 100 is made of a plastic material with high hardness, and the material of the cover 200 is made of polydimethylsilicon (PDMS) which is relatively low in hardness and porous. However, the present invention is not limited to this. The substrate 100 may also be made of a material with low hardness, and the cover 200 is made of a material with high hardness. It is worth noting that, in order to mass-produce the present invention, in addition to selecting the materials of the substrate 100 and the cover 200, they must be applicable to the technology of mold injection. In addition, in order to prevent the substrates 100 and vacuum that have been combined and evacuated, Due to the collision of the cover body 200 during the transportation process, the substrate 100 and the cover body 200 cannot be closely adhered to maintain a vacuum state. The present invention specifically designs the second surface when the first surface 101 and the second surface 201 are adhered and assembled to each other. The edges of 201 are located inside the edges of the first surface 101 (as shown in FIG. 1). Because the first surface 101 and the second surface 201 have a seam when they are bonded to each other, in mass production, the automated equipment only needs to use a sealant to close the seam, and then a vacuum can be applied to the injection port to make the micro A vacuum state is formed inside the flow channel. In the embodiment of the present invention, the detection device 10 after the completion of the evacuation procedure is packaged in a vacuum packaging bag.

In another embodiment of the present invention, the recessed portion 102, the micro-sensing wafer 300, the micro-channel structure 400, and the quantitative control structure 403 are disposed on the substrate 100. In yet another embodiment, the recessed portion 102, the micro-sensing The measurement chip 300, the microchannel structure 400, and the quantitative control structure 403 may be disposed on the cover 200. In another preferred embodiment of the present invention, a plurality of micro-channels 401, a plurality of recesses 102, a plurality of micro-sensing wafers 300, and a plurality of quantitative control structures 403 may be disposed on the substrate 100, via a special substrate 100 In combination with the configuration of the cover body 200, a single detection device 10 can process multiple test samples simultaneously, which is not only efficient but also saves time and cost. In the embodiment, a pre-processing portion 404 (as shown in FIG. 8) is provided between the recessed portion 102 and the injection port 402, which is suitable for separating the sample 600 or mixing with other reagents. Prior to the actual detection of some special specimens 600, impurities in the original specimens 600 must be removed or mixed with other substances in advance, and the pre-processing section 404 of the present invention has the function of providing the aforementioned separation and mixing.

It is worth noting that the detection device 10 of the present invention utilizes a vacuum method, and one of the substrate 100 and the cover 200 is made of a porous material, so that the internal microchannels 401, the recesses 102, and the capacity control grooves 405 are used. A negative pressure is generated relative to the external atmospheric pressure. When the specimen 600 is placed at the injection port 402, the specimen 600 can flow through the depression 102, micro The sensing chip 300 to the capacity control slot 405 completes the test, and it is not necessary to use a power element such as a pump or a valve to provide additional power to drive the sample 600 during the test. FIG. 9 is an experimental record table of the detection device 10 according to an embodiment of the present invention. Please refer to FIG. 9. In the test sample 1, because the detection device 10 has not been evacuated first, although the sample 600 is dropped to the injection port 402 After one day, the specimen 600 still cannot flow to the recessed portion 102. In the test samples 2 to 6, after the test device 10 was vacuumed for 6 minutes and 30 seconds to 7 minutes, the specimen 600 took about 14 At 17 clocks to the entrance of the micro-sensing chip 300. Therefore, it is known from the experiment that the flow pattern of the specimen 600 in the detection device 10 is consistent and repeatable after the detection device 10 of the present invention undergoes a vacuum treatment for a specific time.

Referring to FIG. 10, in another embodiment, a plurality of terminals of the detection device 10 of the present invention may be formed on a terminal carrier plate 500, and the terminal carrier plate 500 is formed on the first surface 101 of the substrate 100 and protrudes. On one side of the substrate 100. The micro-sensing wafer 300 in this embodiment may be similar to the foregoing embodiment; the cover 200, the capacity control groove 405, the injection port 402, the recessed portion 102, and the microchannel 401 may also be similar to the foregoing embodiment.

Please refer to FIG. 11A and FIG. 11B. The detection device 10 of the foregoing embodiments of the present invention may further include: a first-rate choke channel 408 formed in the micro-channel 401; and a reaction tank 409 communicating with the micro-sensing chip 300, wherein A flow resistance flow channel 408 is formed between the capacity control tank 405 and the reaction tank 409 to further delay the flow time of the sample 600 to the quantitative control structure 403, increase the reaction time between the sample 600 and the micro-sensing wafer 300, and thereby improve the detection time. Accuracy. Since the specimen 600 contains the specimen fluid and the target substance to be suspended in the specimen fluid, there must be enough target substance to be deposited on the micro-sensing wafer 300 to make the most accurate analysis. The flow resistance channel 408 enables After the specimen 600 fills the reaction tank 409, the time during which the specimen 600 stays in the reaction tank 409 reaches 3 to 7 minutes, and in one embodiment, it can reach about 5 minutes. The effect of the flow blocking flow channel 408 slowing down the flow rate of the specimen 600 can make a certain amount of the specimen 600 fully react with the micro-sensing wafer 300. When the cavity corresponding to the reaction tank 409 is filled with the specimen 600, the specimen 600 is replaced by The flow resistance flow path 408 continues to flow to the capacity control tank 405, but the volume of the flow resistance flow path 408 is relatively small compared to the reaction tank 40. Therefore, the sample 600 in the reaction tank 409 before the sample 600 flows to the capacity control tank 405 It can be regarded as a stationary state, that is, within a unit time, the micro-sensing wafer 300 reacts with a certain amount of the specimen 600, so the volume of the cavity in which the reaction tank 409 is combined with the substrate 100 can be used as a unit for a certain amount of analysis. The flow resistance flow channel 408 may have a zigzag pattern, such as in the form of more zigzag times, short zigzag amplitude and narrow channel width in FIG. 11A, or in FIG. form.

Please refer to FIG. 12 to FIG. 16, which show that the recessed portion 102 of the substrate 100 of the detection device 10 of the present invention may have different changes. As shown in FIG. 12, the flat bottom 103 of the recessed portion 102 is closer to the injection port 401 than the slope 104. As shown in FIG. 13, the recessed portion 102 may only have a slope 104 without a flat bottom, and the depth of the slope 104 is deeper as it moves away from the injection port 401. As shown in FIG. 14, the recessed portion 102 may have only the slope 104 without a flat bottom. The depth of the slope 104 is further away from the injection. The shallower the mouth 401. As shown in FIG. 15, the width of the recessed portion 102 can be narrower as it moves away from the injection port 401. As shown in FIG. 16, the width of the recessed portion 102 can be wider as it moves away from the injection port 401.

In summary, the detection device 10 of the present invention can separate the heavier molecules from the lighter molecules in the specimen 600 by the design of the recessed portion 102. Because a centrifugal separator is not required and the sample 600 can be directly separated, the detection device 10 of the present invention has the green environmental protection concept of convenience and energy saving in use. When the detection device 10 of the present invention is electrically connected to an external device, the separated specimen 600 can upload the detection signal to the external device at the same time as the detection, so that researchers can conduct subsequent related research and analysis. The detection device 10 of the invention also has the advantages of fast detection and simple operation.

Please refer to the top perspective view of FIG. 17, which shows another embodiment of the detection device of the present invention. The detection device 20 of the present invention includes: a substrate 700 including a recess 702, a microchannel 704, and at least one reaction tank may include, for example, a first reaction tank 706, a second reaction tank 708, a third reaction tank 710, and A fourth reaction tank 712, in which the first to fourth reaction tanks are communicated by microchannels 704; a cover 900 includes a plurality of injection ports such as a first injection port 810, a second injection port 812, and a microchannel 802 804, a first sensing cavity 814, and at least one sensing cavity may include, for example, a second sensing cavity 816, a third sensing cavity 818, a fourth sensing cavity 820, and a certain amount of control structure 803; The micro-sensing wafer 900 is embedded in the substrate 700; and a terminal carrier 720 is connected to the micro-sensing wafer 900 and a plurality of terminals 721 are formed on the surface. The microchannel 802 may include a first portion located between the first injection port 810 and the quantitative control structure 803 and a second portion substantially parallel to the first portion and extending from the quantitative control structure 803 toward the second injection port 812, wherein the microchannel The first part and the second part of 802 are connected to two different positions of the quantitative control structure 803. The cover 800 may further include a first-class choke channel 808 formed on the first portion of the micro-channel 802. As previously stated As can be seen from the examples, the quantitative control structure 803 may be formed on the substrate 700 in addition to the cover 800.

The material of the substrate 700 may be similar to the foregoing embodiment, which is a relatively hard plastic or hydrophilic material, and the material of the cover 800 may be similar to that of the foregoing embodiment, which is a soft PDMS or other hydrophobic material, meaning the cover. 800 is more hydrophobic than substrate 700. After the cover 800 covers the substrate 700, the first portion of the microfluidic channel 802 is connected to the recessed portion 702 of the substrate 700 and one of the first sensing regions of the microsensor wafer 900, the first reaction tank 706, and the second reaction tank 708, the third reaction tank 710, and the fourth reaction tank 712 are closed by the cover 800 and are respectively connected by the branches of the second part of the microchannel 802. The quantitative control structure 803 of the cover 800 and the substrate 100 form a Capacity control slot 805. After the cover 800 covers the substrate 700, the first sensing cavity 814, the second sensing cavity 816, the third sensing cavity 818, and the fourth sensing cavity 820 are respectively covered on one of the micro-sensing wafers 900 and the second. The four different sensing parts of the sensing area, the first sensing cavity 814, the second sensing cavity 816, the third sensing cavity 818, and the fourth sensing cavity 820 are connected to the first through a microfluidic channel 804, respectively. The reaction tank 706, the second reaction tank 708, the third reaction tank 710, and the fourth reaction tank 712. The first portion of the microfluidic channel 802 is selectively widened at the corresponding micro-sensing wafer 900. The micro-channels 802, 804, 704 and the capacity control slot 805 can be regarded as a micro-channel system. Although the detection device 20 of this embodiment has four reaction tanks and four sensing cavities, in other embodiments, the detection device 20 may have only one reaction tank and one sensing cavity. Although the detection device 20 of this embodiment has two sensing areas, it may have only one sensing area in other embodiments. Because the cover 800 is hydrophobic and the substrate 700 is hydrophilic, when a sample is passed into the microfluidic channel 802 through the first injection port 810, the sample is automatically held by one side of the substrate 700 in the microfluidic channel 802. The sample will not be full until the substrate 700 absorbs the sample and exceeds its saturation value for hydrophilicity. Therefore, when the sample flows to a groove such as the recess 702 or the capacity control groove 805, it will also be full. Behind the tank It then flows out from the microfluidic channel 802 and flows to the next tank.

The detection device 20 of this embodiment can be applied to drug resistance detection, as follows:

Step 1: The first reaction tank 706, the second reaction tank 708, the third reaction tank 710, and the fourth reaction tank 712 have been pre-filled with four kinds of antibiotics (not shown) against a bacterium, and are attached to the form of a patch The bottom of each tank.

Step 2: A sample (not shown) is dropped into the first injection port 810, and the sample enters the recessed portion 702 through the first part of the microchannel 802 to filter impurities. According to the foregoing embodiment, it can be known that the vacuum state of the microchannel 802 of the detection device 20 can cause the specimen to automatically flow in the direction of the capacity control groove 805. If necessary, the lid body 800 can press the capacity control groove 805 to deform to form a negative pressure to drive the inspection Body flow.

Step 3: After the recessed portion 702 is filled, the filtered specimen reaches the first sensing area of the micro-sensing wafer 900 through the first part of the microfluidic channel 802. At this time, the micro-sensing wafer 900 first determines whether the specimen is Bacteria to be tested for resistance.

Step 4: Drop the culture solution through the second injection port 812. Since the first reaction tank 706, the second reaction tank 708, the third reaction tank 710, and the fourth reaction tank 712 are connected by the microfluidic channel 704, the communication can be used. The tube principle controls the liquid surface height of the four reaction tanks to 95% of the height of the reaction tanks.

Step 5: The filtered specimen is injected into the capacity control tank 805 after preliminary interpretation by the micro-sensing wafer 900. After the filtered specimen is collected in the capacity control tank 805, the filtered specimen starts to pass through the microfluid. The four branches of the second part of the channel 802 are respectively injected into the first reaction tank 706, the second reaction tank 708, the third reaction tank 710 and the fourth reaction tank 71. After the four reaction tanks are filled, the filtered The specimen then fills the first sensing cavity 814 and the second via the microfluidic channel 804 The sensing cavity 816, the third sensing cavity 818, and the fourth sensing cavity 82.

Step 6: Read the four electrical signals from the four sensing sections of the second sensing area of the micro-sensing chip 900.

Step 7: After culturing the bacteria in the first reaction tank 706, the second reaction tank 708, the third reaction tank 710, and the fourth reaction tank 71 for about half an hour, read the fourth of the second sensing area of the micro-sensing wafer 900 again. Four signals from each sensing unit.

Step 8: Compare the electrical signals of Step 6 and Step 7, if there is a significant change, you can judge the resistance of the corresponding antibiotic. For example, if the electric signal of the micro-sensing chip 900 corresponding to the first sensing cavity 814 in Step 7 is increased compared to Step 6, it means that the bacteria has resistance to the antibiotic in the first reaction tank 706, and if the micro-sensing chip 900 corresponds to Step 7 The electrical signal of the first sensing cavity 814 is not significantly increased compared to Step 6, which indicates that the antibiotic in the first reaction tank 706 has an effect of inhibiting the growth of the bacteria.

The detection device 20 of this embodiment can also be applied to detect exosome in blood, as follows:

Step 1: Drop the blood with extracellular chromosome (not shown) through the first injection port 810. After the blood enters the depression 702 through the first part of the microfluidic channel, the blood cells are intercepted by the depression 702, and the plasma continues to The micro-sensing wafer 900 flows. According to the foregoing embodiment, it can be known that the vacuum state of the microchannel 802 of the detection device 20 can cause the plasma to automatically flow in the direction of the volume control tank 805. If necessary, the cover 800 can press the volume control tank 805 to form a negative pressure to drive the flow of plasma.

Step 2: The lysis buffer is dripped from the second injection port 812. Since the first reaction tank 706, the second reaction tank 708, the third reaction tank 710, and the fourth reaction tank 712 are used by the microfluidic channel 704 Connected, based on the principle of the connecting tube, the liquid level of the four reaction tanks is controlled At 95% reaction tank height. Because the detection device 20 does not need to determine whether the foreign body is a preset type when it is used to detect foreign bodies in the blood, the micro-sensing chip 900 of the detection device 20 may only have a sensing area, a reaction tank, and a micro-connector. One of the sensing wafers 900 senses a cavity.

Step 3: After the volume control tank 805 collects plasma from the first part of the microchannel 802, the plasma is injected into the first reaction tank 706 and the second reaction tank 708 from the four branches of the second part of the microchannel 803. The third reaction tank 710 and the fourth reaction tank 712 react with the cell decomposition solution, which can decompose the cell wall of the exosomes to expose the proteins of the exosomes. When the four reaction tanks are filled, the reacted plasma fills the first sensing cavity 814, the second sensing cavity 816, the third sensing cavity 818, and the fourth sensing cavity 82 via the microfluidic channel 804. .

Step 4: Read the four electrical signals of the four sensing sections of the second sensing area of the micro-sensing chip 900.

Step 5: After the outside of the four reaction tanks starts to react for about half an hour, read the four electrical signals of the four sensing sections of the second sensing area of the micro sensing chip 900 again.

Step 6: Compare the electrical signals of Step 5 and Step 4 and estimate the concentration of the foreign body from the change of the electrical signals of the two steps.

Please refer to FIG. 18, which is a top perspective view showing another embodiment of the detection device of the present invention. The detection device 30 includes: a substrate 700 including a depression 702 and a decomposition tank 760; a cover 800 including a micro-channel 802, a sensing cavity 809, a first injection port 810, and a second injection port 812; A micro-sensing wafer 900 is embedded in the substrate 700 and has a sensing area communicated by the sensing cavity 809. A heating element 750 includes a heating wafer 751 and a resistance wire 752 meanderingly formed on the heating wafer 751; And a terminal carrier board 720 is connected to the micro-sensing chip 900 and a plurality of terminals 721 are formed on the surface. The recessed portion 702 may be the recessed portion of the foregoing embodiments, for example The different forms of Figures 12 to 16, and / or arrayed micro pillars (not shown) or other disclosed structures can be added to the specimen to increase the suspension of the specimen. Probability of sedimentation. The resistance line 752 of the heating element 750 has two conductive ends 752a and 752b respectively electrically connected to two contacts of a power supply (not shown). The temperature of the resistance line 752 will rise and heat the microchannel 802 above. The heating chip 751 can regulate the temperature of the resistance wire 752. The detection device 30 may include a capacity control slot 805 whose quantitative control structure may be formed in the cover 800 or the substrate 700; and a first-rate choke channel 808 formed in the micro flow channel 802 and located between the capacity control slot 805 and the sensing cavity 809 between.

The detection device 30 of this embodiment can be applied to the detection of DNA extracted by Polymerase Chain Reaction (PCR). A specimen having DNA, such as blood (not shown), can be injected into the decomposition tank 760 through the first injection port 810 A cell decomposition solution can be injected through the second injection port 812. After the cell decomposition solution fills the decomposition tank 760, it can flow with the sample from the microchannel 802 to the recess 702, and the sample can be filtered in the recess 702, and then flows from the microchannel 802 to the heating element 750 to be heated. The lane 802 is zigzag on the heating element 750 and its zigzag direction is approximately perpendicular to the zigzag direction of the resistance line 752, so that the specimen can be uniformly heated. After the specimen is repeatedly heated by the heating element 750 between 95 ° C and 65 ° C for many times, the DNA contained therein is copied in large quantities, and then flows from the microchannel 802 to the sensing cavity 809 and the microsensing wafer 900 In response, the micro-sensing chip 900 may also have a plurality of sensing sections to perform different tests on DNA. As in the previous embodiment, the flow blocking flow channel 808 and the volume control groove 805 enable the detection device 30 to analyze the sample quantitatively.

Although the present invention is disclosed as above by way of example, it is not intended to limit the scope of the present invention. Any person skilled in the art can do some things without departing from the spirit of the present invention. Changes and retouching, so the protection scope of the present invention shall be determined by the scope of the appended patent application.

10‧‧‧Detection device

100‧‧‧ substrate

101‧‧‧first surface

102‧‧‧ Depression

200‧‧‧ Cover

300‧‧‧Micro Sensing Chip

401‧‧‧microfluidic channel

402‧‧‧Injection port

405‧‧‧Capacity control slot

501‧‧‧Plural terminals

Claims (29)

A detection device includes: a substrate having a first surface, the first surface having a recessed portion, the recessed portion including a bottom and a slope, the bottom being embedded in the substrate, and the slope connecting the first surface and The bottom is disposed at one end of the recessed portion; a cover has a second surface facing the first surface, a micro-sensing wafer is embedded in the substrate, and a micro-channel structure is embedded in the second surface Wherein, after the cover body covers the substrate, the first surface and the second surface are closely adhered to each other, so that the microfluidic channel structure and the first surface are combined into a microfluidic channel including at least one injection port and a capacity control groove Control the flow rate of a specimen in the microfluidic channel, the specimen enters the depression from the injection port through the microfluidic channel, and the specimen is separated into a lower layer liquid and an upper layer liquid in the depression portion, and the lower layer liquid stays At the bottom, the upper layer fluid flows from the depression to the micro-sensing chip; wherein the micro-channel includes a first-rate blocking channel and a reaction tank, the reaction tank is connected to the micro-sensing chip, and the flow is blocked by the flow. Road formation Between the groove and the capacity control of the reaction tank; wherein the substrate and at least one of the cover comprises a porous material, and the cover of the substrate compared to high hydrophobicity. The detection device according to item 1 of the scope of the patent application, wherein the specimen is blood, the lower fluid is blood cells, and the upper fluid is plasma. The detection device according to item 1 of the scope of patent application, wherein the slope is disposed in the recessed portion near one end of the injection port. The detection device according to item 1 of the patent application scope, wherein the micro-sensing wafer has at least one detection structure, and the detection structure energizes biological particles or biopolymers in the sample. The detection device according to item 1 of the scope of patent application, further comprising a plurality of terminals, which are disposed on the substrate and connected to the micro-sensing chip, and the terminals can be coupled to a reading device. The detection device according to item 6 of the scope of patent application, wherein the terminals are connected to the micro-sensing chip in a wire bonding manner. The detection device according to item 5 of the scope of patent application, wherein the detection structure is a resistance type, a capacitance type, or a transistor type, or an electrochemical sensor based on a nanometer sensing material, a nanometer material After the functionalization of a biopolymer, the biopolymer is selected from the group consisting of antibodies, aptamers or sugar molecules or enzymes, or the detection structure includes non-nanometer electrochemical or photoelectric sensors. The detection device according to item 8 of the scope of patent application, wherein the nanometer sensing material is selected from the group consisting of nanometer carbon tubes, graphene, reduced graphene oxide (rGO), and graphene Graphene oxide (GO), nanoribbon graphene, nano silicon wire, nano InP wire, nano GaN wire, nano semiconductor wire, or nano semiconductor film. The detection device according to item 1 of the scope of patent application, wherein the material of the substrate is polymethylmethacrylate (PMMA), polyethylene terephthalate Ester (polyethylene terephthalate, PET), polycarbonate (PC), polydimethylsilicon (PDMS), porous silicone, rubber, plastic or glass. The detection device according to item 1 of the scope of patent application, wherein the material of the cover body is polymethylmethacrylate (PMMA), polyethylene terephthalate (PET), and polycarbonate (polycarbonate, PC), polydimethylsilicon (PDMS), porous silicone, rubber or plastic. The detection device according to item 1 of the scope of patent application, wherein a pretreatment section is provided between the recessed portion and the injection port, which is suitable for separating the sample or mixing with other reagents. The detection device according to item 1 of the scope of patent application, wherein when the second surface is assembled with the first surface, an edge of the second surface is located inside the edge of the first surface. The detection device according to item 1 of the scope of patent application, wherein the bottom and slope of the recessed portion are coated with calcium chloride or a mixture containing thrombin, fibrinogen, and calcium ion To form a fibrin mesh. The detection device according to item 1 of the scope of the patent application, wherein the arrayed micro-pillars are formed at the interface between the recessed portion and the microfluidic channel, and the spacing between the micro-pillars is less than 3 microns, or An array of micro-pillars arranged at the junction is spaced between 10 to 100 microns, and is formed by matching a plurality of micro-spheres larger than the spacing between the micro-pillars. Multiple voids to intercept suspended matter in the specimen. The detection device according to item 1 of the patent application scope, wherein the microfluidic channel is located in an area between the microsensing wafer and the recessed portion, and a cross-sectional area of the channel in the microfluidic channel is reduced. The detection device according to item 1 of the scope of patent application, wherein the slope is disposed in one end of the recessed portion away from the injection port. The detection device according to item 18 of the scope of patent application, wherein the flow resistance flow path is in a zigzag pattern. The detection device according to item 1 of the scope of patent application, wherein the width of the recessed portion is gradually widened away from the injection port or narrowed away from the injection port in an upper view. A detection device includes: a substrate having a first surface including a recessed portion and at least one reaction tank; a cover body having a second surface covering the first surface of the cover body; and a micro-sensing chip embedded The substrate includes at least one sensing area; a first injection port; a second injection port; and a micro-channel system formed between the substrate and the cover, including a first micro-channel formed on the substrate. The first surface communicates with the reaction tanks and the second injection port, a second microchannel is formed on the second surface of the cover body, and a capacity control tank is formed on the second surface of the cover body and communicates. Connecting the reaction grooves to the second microfluidic channel and a third microfluidic channel to at least one of the sensing regions of the micro-sensing chip; The second microfluidic channel includes a first portion and a second portion. The first portion extends from the first injection port to the capacity control groove and communicates with the recessed portion. The second portion is formed by The capacity control tank extends to the at least one reaction tank; at least one of the substrate and the cover includes a porous material, and the cover is more hydrophobic than the substrate. The detection device according to item 19 of the scope of patent application, wherein the micro-fluid channel system, the at least one reaction tank and the recessed portion are in a vacuum state before detection. The detection device according to item 19 of the scope of patent application, wherein the cover body can be deformed by being pressed at a position corresponding to the capacity control groove. The detection device according to item 19 of the scope of patent application, further comprising a first-class choke channel formed in the first part of the second micro-channel. The detection device described in item 19 of the patent application scope further includes a terminal carrier board electrically connected to the micro-sensing chip and having a plurality of terminals. The detection device according to item 19 of the scope of patent application, wherein the sensing area includes a first sensing area connected by the first part of the second microfluidic channel and a second sensing area is connected by the third micro area. The flow channel is connected. A detection device includes: a substrate having a first surface including a recessed portion and a decomposition groove; a cover body having a second surface covering the first surface of the cover body; and a micro-sensing chip embedded in the The substrate includes a sensing area; a microchannel is formed between the substrate and the cover, and communicates with the recessed portion and the The sensing area of the micro-sensing wafer; a heating element is formed under a part of the micro-fluidic channel; one of the decomposition solution contains a DNA-containing sample that fills the decomposition tank and the depression and flows through the micro-fluidic channel Above the heating element, a large number of copies are circulated and heated by the heating element, and then flowed to the sensing area of the micro-sensing wafer through the micro-fluidic channel; at least one of the substrate and the cover includes porosity The material and the cover are more hydrophobic than the substrate. The detection device according to item 25 of the scope of patent application, wherein the arrayed micro-pillars are formed at the boundary between the recessed portion and the microchannel, and the spacing between the micropillars is less than 3 micrometers, or between the recessed portion and the microchannel. An array of micro-pillars arranged at the junction is spaced between 10 to 100 micrometers, and a plurality of microspheres are larger than the space between the micro-pillars to form a plurality of gaps to intercept suspended matter in the specimen. The detection device according to item 25 of the patent application scope further includes a first injection port for injecting the specimen and a second injection port for injecting the decomposition liquid. The detection device described in item 25 of the scope of patent application, further includes a first-class choke channel, a capacity control slot, and a sensing cavity, wherein the sensing cavity is connected to the micro flow channel and to the sensing area, the A flow resistance flow channel is formed in the micro flow channel and is located between the capacity control slot and the sensing cavity. The detection device according to item 25 of the scope of patent application, wherein the heating component includes a heating wafer and a resistance line located on the heating wafer in a zigzag shape, and a portion of the microfluidic channel on the resistance line is also zigzag shape. And its zigzag direction is perpendicular to the zigzag direction of the resistance line.