CN103387935A - Microfluidic array chip for cell capture - Google Patents

Abstract

The invention relates to a microarray structure chip utilizing the difference in diameter of different types of cells, adopting a microfluidic pipeline, and etching microcolumn arrays with different pitches and sizes in the pipeline. The chip structure is composed of a silicon Si material base plate and a The soda-lime glass upper cover is bonded up and down. According to the different spacing of the microstructure of the column, it can be divided into two series: variable spacing and fixed spacing; according to the size of the microstructure, it can be divided into various types: ranging from 2-30μm. It is characterized in that when the liquid fluid containing different types of cells flows through the microstructure of the chip, when passing through the microarrays of pillars with different pitches and diameters, the cells of different types (different diameters) will be covered by different micropillar array structures. Capture, stay on the cylinder arrays with different microstructures in the pipeline, and use a microscope to observe and analyze the capture results. The invention has the characteristics of micro volume, flexible and customizable structure, and no moving parts, etc., and can be widely used in the fields of capture and analysis of various cancer cells in blood, and uses different types of chips to comprehensively analyze the types and quantities of cells in the blood of patients and calculation judgment.

Description

Cell capture microfluidic array chip

technical field

The invention belongs to the field of biological cell detection in a fluid, in particular to a design of a cell capture chip utilizing a micro-column array structure distributed in a micron-scale micro-flow tube. the

Background technique

The microfluidic chip originally originated in the field of analytical chemistry. It is a kind of micro-cavity, micro-pipeline network structure and other functional units made on a substrate of several square centimeters by using fine processing technology to realize the preparation and injection of micro-sample. , reaction, separation and detection in one of the rapid, efficient, low-cost micro-analysis experimental device. the

The use of microfluidic chips for blood sample analysis is a scientific research hotspot that has emerged in recent years. For example, a team led by Mehmet Toner, a professor of biomedical engineering at Harvard Medical School, designed a small device in 2011 that could detect cancer cells in blood samples. The microfluidic device is the size of a coin and can detect a variety of viruses, including HIV. The core structure of the microfluidic device is a microcolumn made of carbon nanotubes with a diameter of 30 microns, which can capture any cancer cells or other small substances flowing through the device. This microfluidic device holds promise for detecting cancer cells that have spread through blood. the

Domestic research in this field is still in its infancy. The invention utilizes silicon materials and soda-lime glass materials to make a cheap cell capture microarray chip to capture various types of cells in blood including cancer cells, and can be widely used in various processes involving the analysis of blood cell composition. The field of disease analysis. the

Contents of the invention

The present invention proposes a cell capture microfluidic array chip structure, which etches a microarray structure of micro-column arrays with different pitches and sizes in the pipeline. combined to form. Utilizing the difference in diameter of different types of cells in different types of blood cells, after flowing through the microarray capture area with different column diameters and spacings, they will be layered and trapped in different positions of the chip, and the number of cells trapped in different positions can be analyzed to accurately judge The content of various cells in blood can be widely used in the capture and analysis of various cancer cells in blood, hospital research, pharmaceutical clinical trials, tumor diagnosis, personalized treatment and other fields. the

In order to achieve the above purpose, the technical solution of the present invention is: a cell capture microfluidic array chip, which can be divided into two series according to the different spacing of the column microstructure: variable spacing and fixed spacing; according to the size of the microstructure, it can be divided into two series: There are many types: ranging from 2-30μm. It is characterized in that a series of microcolumns are made of silicon wafers, and antibodies are coated on the surface. When the liquid fluid containing different types of cells flows through the microstructure of the chip, when passing through the microarrays of cylinders with different pitches and diameters, different Cells with different diameters (types) will be captured by different micro-pillar array structures, and stay on the micro-pillar arrays with different microstructures in the pipeline, and the capture results will be analyzed by microscope observation. the

Features and effects of the present invention are as follows:

1. The present invention has the characteristics of micro volume, flexible and customizable structure, and no moving parts. the

The size of the cell capture microfluidic array chip is about 31mm×24mm×1.5mm. It is made of a common silicon chip and a common soda-lime glass slide. It is small in size and easy to store and transport in batches. the

The arrangement of micro-pillar arrays in the chip can be flexibly customized according to different detection objects. For example, to detect the content ratio of all types of cells in blood, arrays with different diameters and spacing between columns can be arranged in sequence along the direction of blood flow in the microchannel. ; If detecting cells of a single size, an array with a suitable spacing can be arranged in the microchannel according to a certain size, which is used to capture all the target cells flowing through the channel as much as possible and filter cells of other sizes. the

There are no moving parts in the chip, and all the micro-pillar arrays are made by etching the surface of the silicon base, which is integrated with the base, and it is not easy to fall off once the production is completed; the connection between the silicon chip and the glass slide is cured and bonded by a strong ultraviolet light source , to ensure the tightness of the pipeline. The solidified chip structure makes the use process simple and efficient, and the operation is simple. the

2. Use different types of chips to conduct comprehensive analysis and calculation and judgment on the type and quantity of blood cells. The micropillar array is used to capture target cells, and the number of cells captured by the array and the ratio of different types of cells are directly observed through a microscope, which can effectively reflect the blood cell components. the

3. Utilizing mature low-cost consumer electronics manufacturing technology, large-scale and low-cost production of high-end medical products can be rapidly realized. The preparation materials involved in the present invention mainly include silicon wafers, soda-lime glass, and rubber fine catheters. The preparation methods mainly involved are mature dry and wet etching in micro-processing technology, strong ultraviolet bonding and other processes, and the materials are relatively cheap. , low manufacturing cost. the

Description of drawings

Fig. 1 Schematic diagram of chip bonding structure, in which (1) is a silicon substrate, (2) is a soda-lime glass upper cover, (3) is a cell capture micro-pillar array, (4, 5) is an array with capture columns (6) is the inlet and outlet of the drainage tube, and (7) is the drainage rubber tube. the

Fig. 2 Schematic diagram of the structure of the chip silicon matrix substrate (chip silicon substrate (1)), wherein (3) is a cell capture micropillar array, and (4) is a microfluidic channel on a 4-60 μm deep substrate. the

Fig. 3 Schematic diagram of the structure of the chip soda-lime glass matrix upper cover (chip soda-lime glass upper cover (2)), wherein (5) is a 120 μm deep upper cover microfluidic pipeline, and (6) is the inlet and outlet of the drainage tube. the

Figure 4 is a schematic diagram of the arrangement of the micropillar array (cell capture micropillar array (4)), wherein (1) is a silicon microfluidic tube substrate, and (3) is a micropillar array evenly arranged at a certain distance. the

Detailed ways

The present invention is described in detail as follows in conjunction with accompanying drawing:

The overall structure of the chip is shown in Figure 1, including a silicon microfluidic tube substrate (1), a glass upper cover (2), a cell capture micropillar array (3) evenly arranged at a certain distance, and a 4-60 μm deep substrate. On-chip microfluidic conduit (4), 120 μm deep upper cover microfluidic conduit (5), drainage tube inlet and outlet (6), drainage rubber tube (7). the

The structure of the silicon material microfluidic tube substrate (1) is shown in Figure 2, which is divided into silicon wafers, and its size is about 31mm×24mm×0.3mm. the

The structure of the upper cover (2) made of soda-lime glass is shown in Figure 3. It is made of ordinary soda-lime glass, and its size is about 31mm×24mm×1.2mm. the

The structure of cell capture micropillar arrays (3) evenly arranged at a certain interval is shown in Figure 4, which is formed by dry etching of silicon wafers. The schematic diagram of the arrangement of cell capture micropillar arrays (3) shows its structure. basic structural features. The micro-pillar array is arranged in a diamond shape as a whole, and the distance between every two adjacent cylinders is exactly the same. According to the size difference of the captured target cells, it can be divided into 4 kinds of diameters: 2μm, 4μm, 6μm, 10μm, and its height can be divided into 3 categories : 4 μm ~ 60 μm. the

The structure of the microfluidic pipeline (4) on a 4-60 μm deep substrate is shown in Figure 2. It is a multi-μ-shaped structure consisting of 12 straight pipelines (including 2 curved straight lines) and 11 angled pipelines. The width of the pipes is 1mm, and the depth of the pipes matches the array of cell-capturing micropillars (3) uniformly arranged at a certain interval, and can be divided into two series: variable interval and fixed interval. Fixed-pitch series: fixed-pitch 4-10 μm, column diameter 10 μm, depth 8-60 μm; variable-pitch series: every two adjacent straight lines (including curved straight lines) have the same internal spacing, the third straight line has a variable spacing, 12 straight lines The distance between the pipes is 60 μm to 20 μm, the depth of the pipes is 60 μm, and the column diameter can be divided into 4 categories: 2 to 10 μm. the

The structure of the microfluidic channel (5) on the 120 μm deep upper cover is shown in Figure 3, which is completely consistent with the shape and width of the microfluidic channel (4) on the 4-60 μm deep substrate, and only the depth of the channel is different. the

The specific usage of the chip is as follows:

The fixed spacing series is used to comprehensively capture cells of a specific type of diameter. The selection principle is to match the target diameter, that is, the spacing of 10 μm is used to capture cells near the diameter of 10 μm, such as eosinophilic and basic lobulated nuclei (10-16 μm in diameter) , Neutral segmented granulocytes (10-14 μm in diameter); 4 μm apart to capture cells around 4 μm in diameter, such as normal human erythrocytes (about 7 μm in diameter), hypochromic microcytic anemia and simple microcytic anemia cells ( 6.2-6.7 μm in diameter). the

The variable-pitch series is used to analyze blood cell components. The selection principle is based on the actual capture volume requirements. For example, if you only need to analyze whether certain cells are contained, choose a chip with a larger column diameter; if you need to analyze detailed cell components, choose a chip with a smaller column diameter. diameter chips. the

When in use, the blood is sent into the microfluidic pipelines (4, 5) at a low speed by a fixed-speed micropump at one end of the drainage tube inlet and outlet (6), and the blood cells are absorbed by the micropillar array (3) when passing through the micropillar array (3) with the same diameter The trap stays in the gap in the middle of the array. After the blood has completely flowed through the micro-channel (4, 5), the distribution of the captured cells in the micro-channel (4, 5) is observed with a microscope for medical analysis and judgment. the

Claims (8)

1. A cell capture microfluidic array chip, characterized in that, using the difference in diameter of different types of cells in blood of different types of cells, after flowing through the microarray capture areas with different column diameters and spacings, they will be intercepted and separated in layers At different positions of the chip, analyze the number of cells trapped in different positions to accurately determine the content of various cells in the blood. 2. The cell capture microfluidic array chip according to claim 1, the structure includes a silicon material microfluidic tube substrate (1), a glass material upper cover (2), and cell capture microcolumns evenly arranged at a certain distance An array (3), a microfluidic pipeline (4) on a substrate with a depth of 4-60 μm, a microfluidic pipeline (5) with a depth of 120 μm, an inlet and an outlet of a drainage tube (6), and a drainage rubber tube (7). 3. The cell capture microfluidic array chip according to claims 1 and 2, characterized in that it has the characteristics of micro volume, flexible and customizable structure, and no moving parts. The size of the cell capture microfluidic array chip is about 31mm×24mm×1.5mm. It is made of a common silicon wafer and a common slide glass. It is small in size and easy to store and transport in batches. 4. The cell capture microfluidic array chip according to claims 1, 2, and 3, characterized in that the arrangement of micropillar arrays in the chip can be flexibly customized according to different detection objects, such as detecting all types of cells in blood content ratio, arrays with different column diameters and column spacing can be arranged sequentially along the blood flow direction in the microchannel; if cells of a single size are detected, arrays with a suitable spacing can be arranged in the microchannel according to a certain size for Try to completely capture all the target cells flowing through the pipeline and filter cells of other sizes. 5. The cell capture microfluidic array chip according to claims 1, 2, 3, and 4, wherein there are no moving parts in the chip, and all micropillar arrays are made by etching the surface of the silicon base, It is integrated with the base, and it is not easy to fall off once the production is completed; the connection between the silicon wafer and the glass slide is cured and bonded by a strong ultraviolet light source, which ensures the sealing of the pipeline. The solidified chip structure makes the use process simple and efficient, and the operation is simple. 6. The cell capture microfluidic array chip according to claims 1, 2, 3, 4, and 5, characterized in that, using mature low-cost consumer electronics manufacturing technology, large-scale and low-cost high-end medical products can be rapidly realized Production. The preparation materials involved in the present invention mainly include silicon wafers, silica glass, and rubber fine catheters. The preparation methods mainly involved are mature dry and wet etching in microprocessing technology, strong ultraviolet bonding and other processes. The materials are relatively Inexpensive, low manufacturing cost. 7. The cell capture microfluidic array chip according to claims 1, 2, 3, 4, 5, and 6, whose structure is characterized in that the structure of the microfluidic pipeline (4) on a 4-60 μm deep substrate is shown in Figure 2 , into a multi-μ-shaped structure, consisting of 12 straight pipes (including 2 curved straight lines) and 11 angled pipes. The width of the pipes is 1mm, and the depth of the pipes matches the array of cell-capturing micropillars (3) uniformly arranged at a certain interval, and can be divided into two series: variable interval and fixed interval. Fixed spacing series: fixed spacing 10-4μm, column diameter 10μm, depth 60-8μm; variable spacing series: the inner spacing of every two adjacent straight lines (including curved straight lines) is the same, the spacing of the third straight line varies, 12 straight lines The distance between the pipes is 60-20 μm, the depth of the pipes is 60 μm, and the column diameters can be divided into 4 categories: 2-10 μm. 8. The cell capture microfluidic array chip according to claims 1, 2, 3, 4, 5, 6, and 7, its method of use is characterized in that the fixed-pitch series is used to comprehensively capture cells of a specific type of diameter, and the selection principle It matches the target diameter, that is, the spacing of 10 μm is used to capture cells with a diameter of 10 μm; the spacing of 4 μm is used to capture cells with a diameter of 4 μm. The variable-pitch series is used to analyze blood cell components. The selection principle is based on the actual capture volume requirements. For example, if you only need to analyze whether certain cells are contained, choose a chip with a larger column diameter; if you need to analyze detailed cell components, choose a chip with a smaller column diameter. diameter chips. When in use, the blood is sent into the microfluidic pipelines (4, 5) at a low speed by a fixed-speed micropump at one end of the drainage tube inlet and outlet (6), and the blood cells are absorbed by the micropillar array (3) when passing through the micropillar array (3) with the same diameter The trap stays in the gap in the middle of the array. After the blood has completely flowed through the micro-channel (4, 5), the distribution of the captured cells in the micro-channel (4, 5) is observed with a microscope for medical analysis and judgment.

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