CN211402075U - Micro-fluidic automatic separation and intelligent component identification system - Google Patents

Abstract

The utility model discloses a microfluidic automatic sorting and component intelligent identification system. The system includes a droplet chip, a sample loading and storage unit, a particle image detection unit, a particle component identification unit, and an automatic stage shifting device; a particle image detection unit and a plurality of particle groups are sequentially arranged above the sample loading and storage unit. Sub-identification unit; the automatic displacement device of the stage includes a stage and a chip displacement track, the droplet chip is mounted on the stage, and the stage is set on the chip displacement track, and the chip displacement track is respectively connected with the sample loading and storage units, The particle image detection unit and the particle component identification unit are connected; the system of the utility model can realize continuous automatic counting, particle size distribution and image identification, sorting and collection, and intelligent multifunctional analysis and identification of the particles in the micro-solution.

Description

A microfluidic automatic sorting and component intelligent identification system

technical field

The utility model relates to the technical field of particle analysis, in particular to a microfluidic automatic sorting and component intelligent identification system.

Background technique

The requirements for particle control in liquid products are gradually strengthened in the fields of medical, health and environmental protection, such as blood sample analysis, food and pharmaceutical R&D and production quality control, and pure water treatment. Particle detection has also become a vital part of product quality control. Understanding and mastering the particle size distribution, morphological characteristics and structural composition of particles provides direct and powerful ideas and solutions for product process development and process optimization, product quality control and product problem investigation and research in related fields.

Particle size analyzers based on image analysis have been widely reported. Known from US Patent No. US 7064826 B2, a digital optical particle size detection method utilizes reduced magnification to perform pixel point difference analysis on the obtained particle image pixel array, thereby obtaining the detection results of particle size distribution and morphology. A particle detector is known from US Patent No. US 9360410B2 to cover a wide range of particle sizes. Among them, the lower size range and the middle size range are detected based on the dark image area of the particles, and the larger size range of the particles is detected based on the brighter image area of the particles. The particle detector includes a sample flow cell, a dark area light source, a bright area light source, an imaging system, a process system, and a pump system. The liquid sample is driven by a peristaltic pump through the sample flow cell, and the particles in the liquid are captured by the microscope and camera system.

Due to the wide range of application fields of particle detection, there are many fields that require the minimum sample volume to be tested. The sample processing system fabricated with microfluidic chip technology can achieve the goal of micro-quantity and high-throughput detection of liquid samples. It is known from CN 104846400 B that an electrolysis device based on the principle of electrowetting on a dielectric layer obtains droplets containing electrolysis products with specific concentrations and different polarities by controlling the electrolysis process and the droplet splitting process. From CN 104994955 B, droplet manipulation systems are known to manipulate droplets by supplying voltage pulses to individual electrodes in an electrode array to achieve electrowetting.

The current particle detector can only analyze the particle size distribution and morphology of the particles in the liquid, but cannot provide more in-depth information on the structure and composition of the particles. For the morphological analysis of particles, it is still in the preliminary application stage of images. The particle image information obtained by the instrumental analysis cannot be used to accurately judge the properties of the particle, namely endogenous, endogenous and heterologous. This limitation makes particle image analysis often only used as an aid and reference for particle analysis. The only way to know the true structure and origin of particles is to use particle image analysis in conjunction with other particle component identification methods. However, current particle component identification methods rely purely on manual filtration separation of particles in liquids, which is time-consuming and labor-intensive. A trained researcher can only complete the separation and component identification of particles in a liquid sample in 4 hours. Based on the large volume requirement of manual sample operation, the destructiveness to the sample, the unavoidable risk of environmental contamination during sample detection, and the high technical difficulty of separate identification, detection and data analysis, particle component identification is not yet available in the industry. Widespread use.

Utility model content

In order to overcome the deficiencies of the prior art, the purpose of the present invention is to provide a microfluidic automatic sorting and component intelligent identification system. The system has a high degree of automation. It uses a droplet chip made by microfluidic technology and an optical detector for particle image detection and particle component identification, which can realize continuous automatic counting, image and particle size identification of particles in trace solutions, and separation of particles. Selection and collection, as well as multi-functional intelligent analysis and identification of components.

The utility model automatically separates the liquid containing particles into droplets under the traction of the electrowetting technology, and reaches the visible detection window through the digital droplet microfluidic flow path of the droplet chip, and the droplets are arranged on the positive side of the detection window. Detected by an external optical detector above, based on the detected particle image and spectral information, automatic intelligent identification of particle components is realized. The technical solutions of the present utility model are specifically introduced as follows.

A microfluidic automatic sorting and component intelligent identification system, comprising a droplet chip, a sample loading and storage unit, a particle image detection unit, a particle component identification unit and an automatic stage shifting device; sample loading and storage The unit includes a temperature controller and a waste liquid bottle; the automatic displacement device of the stage includes a stage and a chip displacement track, the droplet chip is mounted on the stage, and the stage is arranged on the chip displacement track, and the chip displacement track is respectively and The sample loading and storage unit, the particle image detection unit and the particle component identification unit are connected; the droplet chip includes a bottom plate, an electrode microarray layer, an upper hydrophobic polymer layer, a lower hydrophobic polymer layer and an electrode selector, and at least the bottom of the bottom plate is adhered An electrode selector, an electrode microarray layer is arranged above the bottom plate, a lower hydrophobic polymer layer and an upper hydrophobic polymer layer are arranged above the electrode microarray layer; a sample liquid storage area and a digital solution are arranged between the upper and lower hydrophobic polymer layers The droplet microfluidic flow path, the detection window and the droplet storage area; the sample liquid storage area consists of at least one sample liquid storage chamber; the digital droplet microfluidic flow path includes the main flow path and the branch flow path, the main flow path and each sample storage area. The bottom of the liquid chamber is connected to the liquid outlet, the main flow is connected to the inlet of the detection window, and the outlet of the detection window is connected to each droplet storage chamber through the branch flow path. The distribution of the microfluidic flow path and the distribution of the electrode microarray in the electrode microarray layer Correspondingly, the main flow path mainly realizes the separation of droplets; the droplet storage area includes several droplet storage cells, the outlet of the detection window is divergently connected to each droplet storage cell through the branch flow path, and the main flow path of the inlet of the detection window and the outlet are connected to each other. Electrode microarrays are evenly distributed in the branch flow paths; the particle image detection unit includes a first detector, and the first detector is an image capture device; the particle component identification unit includes a second detector, and the second detector is selected from an ultraviolet spectrometer, an infrared Any one or more of spectrometers, Raman spectrometers, fluorescence spectrometers, microwave spectrometers, electron spin spectrometers or nuclear magnetic resonance spectrometers.

In the utility model, a micro battery is also included, and the micro battery is connected with the electrode selector.

In the present invention, the image capture device is an electron microscope or an optical microscope.

Compared with the prior art, the beneficial effects of the present utility model are:

1. The utility model utilizes digital electrowetting droplet microfluidic technology to process liquid particle samples, which can greatly reduce the consumption of sample volume, and realize the non-destructive analysis of automatic separation of particles in liquid and samples, that is, the analysis of sample particles. Collect and reuse.

2. The utility model combines the microfluidic technology and the particle image detection technology to realize the preliminary automatic sorting of the particles. For example, the common bubbles and silicone oil in the pharmaceutical process can be eliminated in the preliminary sorting.

3. The utility model also adopts the particle component identification technology, which can efficiently complete the component identification of suspected particles on the basis of the preliminary sorting of particles.

4. The utility model adopts the automatic shifting device of the stage, which can realize the fully automatic integrated operation of particle separation and identification.

5. The system of the present invention utilizes the droplet chip of digital microfluidic technology and the integrated multifunctional optical detector, which can realize the continuous automatic counting, particle size distribution and image recognition of protein particles, cells and other particles in the trace solution. , sorting and collection, and multi-functional intelligent analysis and identification of particle components. The system can be applied in any area of process development and quality control where particles are the main component or impurity. For example, the system can be applied to quality control in the production of medicinal liquids, especially for deviation investigation and root cause analysis of particulate impurities. Using the large amount of high-value information generated by the system, it is possible to understand and confirm the reasons for the introduction or formation of particles, and to improve the production process of liquid preparations in a targeted manner, and finally achieve effective control of particles in the production process. In addition, the system can also be applied to formulation and process development, microencapsulation process development, cell clone screening and cell culture process optimization, identification of environmental bacteria and microorganisms, chromatography column packing process detection and quality control, food and beverage Formulation process development and quality control, equipment cleaning process confirmation, and diamond micronization grinding process development and optimization, sewage treatment and pure water process development, and blood sample analysis and other fields.

Description of drawings

FIG. 1 is a schematic structural diagram of the system of the present invention.

FIG. 2 is a schematic diagram of the structure of the droplet chip and the detection of droplets.

FIG. 3 is a schematic diagram of the displacement control of droplets on the droplet chip.

Reference numerals: 1-waste liquid bottle, 2-chip displacement track, 3-waste liquid pipeline, 4-temperature controller, 5-droplet chip, 6-detection window, 7-data processing center, 8-sample loading and storage unit, 9-light source, 10-optical device, 11-first detector, 12-particle image detection unit, 13-second detector, 14,16-particle composition identification unit, 15-nth detector , 17-bottom plate, 18-electrode selector, 19-electrode microarray layer, 20-lower hydrophobic polymer layer, 21-droplet microfluidic flow channel layer, 22-upper hydrophobic polymer layer, 23-liquid to be tested drop, 24-polymer film, 25-sample reservoir, 26-sample reservoir, 27-droplet reservoir, 28-digital droplet microfluidic flow path, 29-droplet reservoir, 30-discard droplets.

Detailed ways

The technical solutions of the present utility model will be described in detail below with reference to the accompanying drawings and embodiments.

As shown in Figures 1 to 3, a microfluidic automatic sorting and component intelligent identification system includes a sample loading and storage unit 8, a droplet chip 5, a particle image detection unit 12, and a particle component identification unit 14. Stage automatic shifting device and data processing center 7 . A particle image detection unit 12 and a particle component identification unit 14 are stacked in sequence directly above the sample loading and storage unit 8; the stage automatic shifting device includes a stage and a chip displacement track 2, and a droplet chip 5 is mounted on the stage , the stage automatic displacement device is connected to the droplet chip 5 and the sample loading and storage unit 8 , the particle image detection unit 12 and the particle component identification unit 14 . The automatic shifting device of the stage can realize two-way transportation. The sample loading and storage unit 8 contains the temperature controller 4 and a waste bottle 1 . When the sample in the droplet chip 5 needs to be heated or cooled and incubated, the stage automatic displacement device transfers the droplet chip 5 to the unit. In order to ensure the analysis of temperature-sensitive biological products, the temperature range of the controllable sample is 2~80℃. After the temperature treatment is completed, the droplet chip 5 is sequentially transferred to the particle image detection unit 12 and the particle component identification units 14 and 16 for detection through the automatic stage shifting device. The particle size range that can be handled by the system of the utility model is wide, and the particle size can be between 0.1 and 1000 μm.

The droplet chip 5 is the sample processing core element of the integrated system of the present invention; the manufacture of the droplet chip 5 is a

An electrode microarray layer 19, an upper hydrophobic polymer layer 22 and a lower hydrophobic polymer layer 20 are sequentially laid on the bottom plate 17; a droplet microfluidic flow path is arranged between the upper hydrophobic polymer layer 22 and the lower hydrophobic polymer layer 20 Layer 21, at least one electrode selector 18 is adhered under the bottom plate 17 of the droplet chip 5. The electrode selector 18 can apply a pulsed voltage to the electrode microarray layer 19, thereby driving the liquid in the droplet microfluidic flow channel layer 21. The droplets are separated and moved by electrowetting. The droplet chip 5 can be made into a film by integral casting of a polymer, or it can be formed by casting the upper and lower layers of the film separately with a polymer, and then adhering to it. The droplet chip 5 can handle a wide range of sample volumes, preferably 1 μL to 1 mL.

The droplet microfluidic flow path layer 21 in the droplet chip 5 is divided into several areas, including a sample liquid storage area 26 , a digital droplet microfluidic flow path 28 , a detection window 6 and a droplet storage area 27 .

The sample liquid storage area 26 is composed of at least one or more (preferably 10) independent 1 mL sample liquid storage chambers 25, which can respectively hold at least one or more same or different biological samples. The sample liquid storage area 26 is a small liquid storage cell without a top cover made of a polymer film (the small cell can be sealed with an additional cover plate, preferably a customized silicone sheet), and the size of the small cell is preferably 10mm×10mm×10mm.

The digital droplet microfluidic flow path 28 is the meridian of the droplet chip 5 and is a droplet manipulation system based on microfluidic electrowetting technology. The droplets are separated from the sample liquid storage chamber 25 , pulled by the droplet manipulation system, flow through the digital droplet microfluidic flow path 28 , and transported to the detection window 6 . The optical detectors 11 , 13 and 15 in the particle image detection unit 12 and the particle component identification unit 14 above 6 detect and identify.

The sample liquid storage chamber 25 to the detection window 6 is a one-way flow path design. The bottom of each sample liquid storage chamber 25 has a main flow path leading to the detection window 6, and the detection window 6 leads to a branch flow path; the bottom of the sample liquid storage chamber 25 has a microchannel. The distribution of the fluidic flow paths corresponds to the distribution of the electrode microarrays in the electrode microarray layer 19. The chip has no special valve design, and the flow of the liquid depends on the electrode selector 18 under the bottom plate 17 to the electrode microarray layer 19. Pulse voltage applied to the electrodes. When the sample liquid storage chamber 25 is filled with liquid and stored liquid, the liquid outlet at the bottom thereof is in a closed state under the action of the pulse voltage. When the sample is detected, the liquid outlet at the bottom is open. Depending on the detection method, the duration of the exit opening is also different. In the continuous liquid flow detection mode, the outlet can be opened for a long time until the liquid in the sample liquid chamber 25 is exhausted; in the droplet detection mode (discontinuous), the opening of the outlet can separate the droplet volume according to the desired size is controlled. The sample reservoir 25 is relatively independent, and there is no risk of cross-contamination. Besides the sample to be tested, the sample liquid storage chamber 25 can also hold a cleaning solution. The cleaning solution can be used for cleaning the microfluidic flow path and the visual detection window 6 when switching between different samples. The cleaning solution may include, but is not limited to, pure water, formulation water, aqueous solutions containing surfactants (Tween 20 or 80), and the like.

The detection window 6 is a visible window, and the preferred diameter of the window is 1 mm in order to realize the detection of a small amount of samples. Below this detection window 6, no electrode microarray is provided. However, electrode microarrays are distributed in the microfluidic flow paths at the inlet and outlet of the detection window 6 to realize the movement of droplets. With the aid of the light source 9 (directly below the chip) and the optics 10 (directly above the detection window), the particles in the droplet are placed in the particle image detection unit 12 and the particle composition identification unit 14 directly above the outside of the detection window 6 The multifunctional optical detectors 11, 13 and 15 are detected. Based on the number and type of particles detected, the droplets can be further divided into smaller droplet units containing only one particle, if necessary. The particle image detection unit 12 includes a light source 9, an optical device 10, and a first detector 11, which is an image capturer (a microscope with automatic focus adjustment), which transmits data based on particle images captured by the microscope to In the processing center 7, the system automatically records the number and size of the particles, and analyzes and judges the suspected properties of the particles; it can identify the silicone oil and air bubbles in the liquid droplets that are common in the production of biological agents, and send them to the waste liquid pipeline 3. In the waste liquid bottle 1 of the external sample loading and storage unit 8, other droplets containing suspicious particles will be transported to the droplet storage chamber 29 of the droplet storage area 27 by the flow path of the droplet manipulation system. A second round of particle composition identification was performed.

The detection window 6 and the droplet storage area 27 are connected by a branch flow path, which is a bidirectional flow path design. The droplet storage area 27 is preferably a square array of 5 cm×5 cm, and the droplet storage area 27 contains enough droplet storage cells 29 to make 12×24=288 2μL storage cells, with a volume of 2μL=2mm ×1mm×1mm, the corresponding number of particles to be tested can be placed and stored. The individual droplet storage cells 29 are independent of each other, and the distance from each other is preferably designed to be 1 mm. In the droplet storage area 27 , the branch flow paths are divergently connected to the respective droplet storage cells 29 . Each droplet storage chamber 29 has an independent number, and the system will mark and number the droplets containing suspicious particles according to the number of the chamber to be inspected. The droplet storage chamber 29 achieves the purpose of particle separation from the sample to be tested, and provides a basis for multifunctional particle identification. The droplets containing suspicious particles temporarily stored in the droplet storage chamber 29 can flow through the digital droplet microfluidic flow path 28 and be retransmitted to the detection window 6 in the droplet chip 5 one by one for detection of particle components; some The special particles can even be transferred back to the sample liquid storage area 26 through the detection window 6, and stored in a closed container after collection, which can be used as a particle standard or for further particle identification and detection. This function satisfies the needs of two or more rounds of detection of suspicious liquid particles. The preferred diameter of the flow path is 1 mm in order to achieve micro-detection of samples. The main features of the liquid chip are small size, controllable droplet volume, controllable droplet flow rate, easy cleaning, not easy to cross-contaminate, and the closed system ensures that the detection process is not susceptible to particle contamination in the external environment.

After the particle image detection of all samples is completed, the droplet chip 5 will be automatically moved to the position of the second detector 13 and then the nth detector 15 , that is, the particle component identification units 14 and 16 . This series of units is connected to a powerful data processing center7. The data processing center 7 contains a rich particle component library, and the built-in intelligent data processing software can automatically compare the particle optical signals collected by the detectors 13 and 15 to the component library, identify and analyze the particle element composition, and then give Accurate judgment of particle properties. The detectors in the particle component identification unit 14 include, but are not limited to, ultraviolet spectrometers, infrared spectrometers, Raman spectrometers, fluorescence spectrometers, microwave spectrometers, electron spin spectrometers, and nuclear magnetic resonance spectrometers. According to the particle identification results, the system automatically compares the particle element database in the system software, and finally generates an identification report of particle attributes.

Claims (3)

1. A micro-fluidic automatic separation and component intelligent identification system is characterized by comprising a liquid drop chip, a sample loading and storing unit, a particle image detection unit, a particle component identification unit and an objective table automatic shifting device; the sample loading and storing unit comprises a temperature controller and a waste liquid bottle, the automatic object stage shifting device comprises an object stage and a chip displacement track, the object stage is loaded with a droplet chip and is arranged on the chip displacement track, and the chip displacement track is respectively connected with the sample loading and storing unit, the particle image detecting unit and the particle component identifying unit; the liquid drop chip comprises a bottom plate, an electrode microarray layer, an upper hydrophobic polymer layer, a lower hydrophobic polymer layer and an electrode selector, wherein at least one electrode selector is adhered below the bottom plate, the electrode microarray layer is arranged above the bottom plate, and the lower hydrophobic polymer layer and the upper hydrophobic polymer layer are arranged above the electrode microarray layer; a sample liquid storage area, a digital liquid drop microfluidic flow path, a detection window and a liquid drop storage area are arranged between the upper hydrophobic polymer layer and the lower hydrophobic polymer layer; the sample liquid storage area is at least composed of a sample liquid storage chamber; the digital liquid drop micro-fluidic flow path comprises a main flow path and branch flow paths, wherein the main flow path is connected with a liquid flow outlet at the bottom of each sample liquid storage chamber, the main flow path is also connected with an inlet of a detection window, the outlet of the detection window is connected with each liquid drop storage chamber through the branch flow paths, the distribution of the micro-fluidic flow path corresponds to the distribution of an electrode microarray in an electrode microarray layer, and the main flow path mainly realizes the separation of liquid drops; the liquid drop storage area comprises a plurality of liquid drop storage small chambers, the outlet of the detection window is divergently connected to each liquid drop storage small chamber through a branch flow path, and electrode microarrays are distributed in the main flow path at the inlet and the branch flow path at the outlet of the detection window; the particle image detection unit comprises a first detector, and the first detector is an image capturer; the particle component identification unit comprises a second detector, and the second detector is selected from one or more of an ultraviolet spectrometer, an infrared spectrometer, a Raman spectrometer, a fluorescence spectrometer, a microwave spectrometer, an electron spin spectrometer or a nuclear magnetic resonance spectrometer.

2. The microfluidic automated sorting and intelligent identity system of components of claim 1, further comprising a micro-battery, wherein the micro-battery is connected to the electrode selector.

3. The microfluidic automated sorting and intelligent identification system of components of claim 1, wherein the image capturer is an electron microscope or an optical microscope.

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