CN103387935A - Microfluidic array chip for cell capture - Google Patents
Microfluidic array chip for cell capture Download PDFInfo
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- CN103387935A CN103387935A CN2012101429049A CN201210142904A CN103387935A CN 103387935 A CN103387935 A CN 103387935A CN 2012101429049 A CN2012101429049 A CN 2012101429049A CN 201210142904 A CN201210142904 A CN 201210142904A CN 103387935 A CN103387935 A CN 103387935A
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Abstract
The invention relates to a microarray-type structure chip adopting micro-flow pipes and taking advantage of diameter difference of different types of cells, and micro column arrays with different intervals and different sizes are etched in the micro-flow pipes. The chip structure is formed by up-down bonding of a silicon Si material negative-film substrate and a soda-lime glass upper cover. According to the different intervals of the column micro structure, the microarray-type structure chip can be divided into two series: variable interval and fixed interval; and according to the different sizes of the column micro structure, the microarray-type structure chip can be divided into a plurality of types: ranging from 2-30 [mu] m. The microarray-type structure chip is characterized in that: when a liquid fluid containing the different types of cells flows through the chip micro structure, and passes through the column microarrays with different intervals and diameters, different types (different sizes) of cells can be captured by different micro column array structures, and retained on the different micro column arrays with different micro structures in the pipes, and the captured results can be observed and analyzed by used of a microscope. The microarray-type structure chip has the characteristics of micro volume, flexible and customizable structure, no moving parts and the like, and can be widely applied to capture and analysis and other fields of various cancer cells in the blood, and comprehensive analysis and measuring and calculating determine of the cell type and quantity in the blood of patients can be conducted through use of different kinds of chips.
Description
Technical field
The invention belongs to biomass cells detection field in fluid, particularly a kind of cell capture chip design that utilizes distribution microcylinder array structure in the micron order microflow channels.
Background technology
Micro-fluidic chip originates from the analytical chemistry field at first, it is a kind of employing retrofit technology, produce microcavity body and microchannel network structure and other functional unit on the substrate of several square centimeters, with realize the preparation of collection micro-example, sample introduction, reaction, separate and be detected on one fast, the micro-analysis experimental installation of efficient, low consumption.
Utilize micro-fluidic chip to carry out the scientific research focus that the blood sample analysis is rising in recent years.For example, research group of the biomedical engineering professor Mei Mitetuona of medical college of Harvard University (Mehmet Toner) leader designed a kind of packaged unit in 2011, can detect the cancer cells in blood sample.This microfluidic device only has one piece of coin-size, the multiple virus that comprises hiv virus can be detected.In microfluidic device, core texture is the miniature column cap of being made by carbon nanotube, and the column cap diameter is 30 microns, can catch cancer cells or other subtitle substances of any device of flowing through.This microfluidic device is expected to detect the cancer cells that spreads by blood.
The research in domestic this field still is in the starting stage.The present invention utilizes silicon materials, soda-lime glass material to make a kind of cell capture micro-array chip of cheapness, comprise that in order to catch blood cancer cells, at interior multiple different sorts cell, can be widely used in the various illness analysis fields that relate to the blood cell compositional analysis.
Summary of the invention
The present invention proposes a kind of cell capture micro-fluidic array chip structure, the microarray type structure of etching different spacing, different big or small microcylinder arrays in pipeline, and this chip structure is by silicon materials egative film substrate and soda-lime glass upper cover bonding and form.Utilize different types of cell dia difference in the different sorts cellular blood, can be held back different positions with chip by layering after the microarray capture region of different column diameters and spacing of flowing through, thereby the cell quantity that the analysis different positions is held back judges the content of various cells in blood exactly, can be widely used in catching and the fields such as analysis, hospital's research, pharmacy clinical trial, diagnosing tumor, personalized treatment of the multiple cancer cells of blood.
For achieving the above object, technical scheme of the present invention is: a kind of cell capture micro-fluidic array chip can be divided into two series according to cylinder microstructure spacing difference: become spacing and determining deviation; Vary in size and can be divided into broad variety according to microstructure: 2-30 μ m does not wait.It is characterized in that, make a series of miniature column caps with silicon chip, antibody is coated in its surface, when the liquid fluid that contains the different sorts cell flows through the chip microstructure, when the cylinder microarray through different spacing and diameter, the cell of different diameter (kind) can be caught by different microcylinder array structures, is trapped on the cylinder array that in pipeline, microstructure is different, utilizes the microscopic examination analysis to catch result.
Characteristics of the present invention and effect are as follows:
1. the present invention has the characteristics such as microbody is long-pending, flexible structure is customizable, movement-less part.
Cell capture micro-fluidic array chip size is 31mm * 24mm * 1.5mm approximately, by a common silicon chip and the common soda-lime glass slide glass of a slice bonding, is formed, and volume is little, is easy to storage carrying in batches.
Arranging of microcylinder array can be according to the different flexible customization of detected object in chip, as detecting the content ratio of all kinds cell in blood, can be in microchannel along blood flow to the different column diameters of arranging successively, the array of different intercolumniations; As detect the cell of single size, can be in microchannel by the arrange array of suitable interval of certain size, the target cell of the pipeline that is used for as far as possible flowing through is fully all caught and is filtered the cell of other sizes.
Movement-less part in chip, all microcylinder arrays are all to be made by the etch silicon susceptor surface, are connected with base, in case the difficult drop-off that completes; Bonding is solidified by strong ultraviolet source in silicon chip and slide glass junction, has guaranteed the stopping property of pipeline.The chip structure that solidifies makes use procedure simply efficient, and operation is succinct.
2. utilize different types of chip to carry out multianalysis and measuring and calculating judgement to blood inner cell kind and quantity.Utilize microcylinder array target acquisition cell, by microscope, directly observe array catch the quantity of cell and the ratio of different sorts cell, can effectively reflect the blood cell component.
3. utilize ripe low-cost consumer electronics manufacturing technology, can realize rapidly high-end medical product on a large scale and low cost production.The material for preparing involved in the present invention mainly comprises silicon chip, soda-lime glass, rubber micro catheter, and the preparation method that relates generally to is the techniques such as dry method, wet etching ripe in micro-processing technology, strong ultraviolet bonding, and material is comparatively cheap, low cost of manufacture.
Description of drawings
Structural representation after Fig. 1 chip bonding, wherein (1) is silicon substrate, (2) be soda-lime glass material upper cover, (3) be that cell catches the microcylinder array, (4,5) are the miniflow pipelines that band catches the post array, (6) be the drainage tube gangway, (7) are the drainage rubber hoses.
Fig. 2 chip silicon matrix egative film (chip silicon substrate (1)) structural representation, wherein (3) are that cell catches the microcylinder array, (4) are miniflow pipelines on 4～60 dark substrates of μ m.
Fig. 3 chip soda-lime glass matrix upper cover (chip soda-lime glass upper cover (2)) structural representation, wherein (5) are the 120 dark upper cover miniflow of μ m pipelines, (6) are the drainage tube gangways.
Fig. 4 microcylinder array (cell catches microcylinder array (4)) arrangement form schematic diagram, wherein (1) is silicon material microflow channels substrate, (3) are the microcylinder arrays that a determining deviation is evenly arranged.
Embodiment
As follows to the detailed description of the invention by reference to the accompanying drawings:
The chip one-piece construction as shown in Figure 1, comprise silicon materials microflow channels substrate (1), glass material upper cover (2), the cell that one determining deviation is evenly arranged catches microcylinder array (3), miniflow pipeline (4) on 4～60 dark substrates of μ m, the 120 dark upper cover miniflow of μ m pipelines (5), drainage tube gangway (6), drainage rubber hose (7).
Silicon materials microflow channels substrate (1) structure as shown in Figure 2, is split to form by silicon wafer, and its size is about 31mm * 24mm * 0.3mm.
Soda-lime glass material upper cover (2) structure as shown in Figure 3, is formed by common soda-lime glass polishing, and its size is about 31mm * 24mm * 1.2mm.
The cell that one determining deviation is evenly arranged catches microcylinder array (3) structure and as shown in Figure 4, by silicon chip, through dry etching, is formed, and cell catches microcylinder array (3) arrangement form schematic diagram and provided its basic structural feature.Microcylinder array integral body assumes diamond in shape and arranges, and every adjacent two cylinder spacings are identical, according to the difference in size of captured target cell, can be divided into 4 kinds of diameters: 2 μ m, 4 μ m, 6 μ m, 10 μ m, its height can be divided into 3 classes: 4 μ m～60 μ m.
On 4～60 dark substrates of μ m, miniflow pipeline (4) structure as shown in Figure 2, becomes many μ type structure, 12 rectilinear duct (containing 2 crooked straight lines) and 11 knuckle pipelines, consists of.Its duct width is 1mm, and the cell that its pipeline depth and a determining deviation are evenly arranged catches microcylinder array (3) and is complementary, and can be divided into 2 series: become spacing and determining deviation.Determining deviation series: determining deviation 4～10 μ m, column diameter 10 μ m, the degree of depth 8～60 μ m; Become spacing series: in every two adjacent straight lines (containing crooked straight line) pipeline, spacing is identical, and the 3rd rectilinear duct spacing changes, and 12 rectilinear duct spacings are 60 μ m～20 μ m, and pipeline depth is 60 μ m, and column diameter can be divided into 4 classes: 2～10 μ m.
The 120 dark upper cover miniflow of μ m pipeline (5) structures as shown in Figure 3, and are consistent with the complete mirror image of miniflow pipeline (4) shape width on 4～60 dark substrates of μ m, and only pipeline depth is different.
The concrete using method of chip is as follows:
Determining deviation series is used for catching the cell of particular types diameter comprehensively, selection principle is to mate with aimed dia, be spacing 10 μ m in order to catch near the cell diameter 10 μ m, for example have a liking for acid, alkaline segmented granulocyte (diameter 10～16 μ m), neutrophilic segmented granulocyte (diameter 10～14 μ m); Spacing 4 μ m are in order to catch near the cell diameter 4 μ m, for example Red Blood Cells of Normal Persons (diameter 7 μ m left and right), low pigment small cell anemia and simple small cell anemia cell (diameter 6.2～6.7 μ m).
Become spacing series and be used for the analyzing blood cellular component, selection principle is according to actual acquisition amount demand and fixed, for example only needs to analyze whether to contain some cells and select the chip of larger column diameter, needs the detailed component of analysis of cells to select the minor diameter chip.
During use, blood is sent in miniflow pipeline (4,5) by determining rotating speed Micropump low speed through drainage tube gangway (6) one ends, and blood cell is caught in the gap of staying in the middle of array by cylinder array (3) during through the similar microcylinder array (3) of diameter.Use captive cell distribution in microscopic microchannel (4,5) after blood flows through microchannel (4,5) fully, carry out the medical analysis judgement.
Claims (8)
1. cell capture micro-fluidic array chip, it is characterized in that, utilize different types of cell dia difference in the different sorts cellular blood, can be held back different positions with chip by layering after the microarray capture region of different column diameters and spacing of flowing through, thereby analyze cell quantity that different positions holds back, judge exactly the content of various cells in blood.
2. cell capture micro-fluidic array chip according to claim 1, structure comprises silicon materials microflow channels substrate (1), glass material upper cover (2), the cell that one determining deviation is evenly arranged catches microcylinder array (3), miniflow pipeline (4) on 4～60 dark substrates of μ m, the 120 dark upper cover miniflow of μ m pipelines (5), drainage tube gangway (6), drainage rubber hose (7).
3. according to claim 1,2 described cell capture micro-fluidic array chips, it is characterized in that having the characteristics such as microbody is long-pending, flexible structure is customizable, movement-less part.Cell capture micro-fluidic array chip size is 31mm * 24mm * 1.5mm approximately, by a common silicon chip and the common slide glass bonding of a slice, is formed, and volume is little, is easy to storage carrying in batches.
4. 2,3 described cell capture micro-fluidic array chips according to claim 1,, it is characterized in that, in chip, arranging of microcylinder array can be according to the different flexible customization of detected object, as detect the content ratio of all kinds cell in blood, can be in microchannel along blood flow to the different column diameters of arranging successively, the array of different intercolumniations; As detect the cell of single size, can be in microchannel by the arrange array of suitable interval of certain size, the target cell of the pipeline that is used for as far as possible flowing through is fully all caught and is filtered the cell of other sizes.
5. 2,3,4 described cell capture micro-fluidic array chips according to claim 1,, it is characterized in that, movement-less part in chip, all microcylinder arrays are all to be made by the etch silicon susceptor surface, be connected with base, in case the difficult drop-off that completes; Bonding is solidified by strong ultraviolet source in silicon chip and slide glass junction, has guaranteed the stopping property of pipeline.The chip structure that solidifies makes use procedure simply efficient, and operation is succinct.
6. according to claim 1,2,3,4,5 described cell capture micro-fluidic array chips, it is characterized in that, utilize ripe low-cost consumer electronics manufacturing technology, can realize rapidly the extensive and low cost production of high-end medical product.The material for preparing involved in the present invention mainly comprises silicon chip, silica glass, rubber micro catheter, and the preparation method that relates generally to is the techniques such as dry method, wet etching ripe in micro-processing technology, strong ultraviolet bonding, and material is comparatively cheap, low cost of manufacture.
7. 2,3,4,5,6 described cell capture micro-fluidic array chips according to claim 1,, its constitutional features is, on 4～60 dark substrates of μ m, miniflow pipeline (4) structure as shown in Figure 2, become many μ type structure, by 12 rectilinear duct (containing 2 crooked straight lines) and 11 knuckle pipelines, formed.Its duct width is 1mm, and the cell that its pipeline depth and a determining deviation are evenly arranged catches microcylinder array (3) and is complementary, and can be divided into 2 series: become spacing and determining deviation.Determining deviation series: determining deviation 10～4 μ m, column diameter 10 μ m, the degree of depth 60～8 μ m; Become spacing series: in every two adjacent straight lines (containing crooked straight line) pipeline, spacing is identical, and the 3rd rectilinear duct spacing changes, and 12 rectilinear duct spacings are followed successively by 60～20 μ m, and pipeline depth is 60 μ m, and column diameter can be divided into 4 classes: 2～10 μ m.
8. 2,3,4,5,6,7 described cell capture micro-fluidic array chips according to claim 1,, its using method is characterised in that, determining deviation series is used for catching the cell of particular types diameter comprehensively, selection principle is and aimed dia coupling, and namely spacing 10 μ m are in order to catch near the cell diameter 10 μ m; Spacing 4 μ m are in order to catch near the cell diameter 4 μ m.Become spacing series and be used for the analyzing blood cellular component, selection principle is according to actual acquisition amount demand and fixed, for example only needs to analyze whether to contain some cells and select the chip of larger column diameter, needs the detailed component of analysis of cells to select the minor diameter chip.During use, blood is sent in miniflow pipeline (4,5) by determining rotating speed Micropump low speed through drainage tube gangway (6) one ends, and blood cell is caught in the gap of staying in the middle of array by cylinder array (3) during through the similar microcylinder array (3) of diameter.Use captive cell distribution in microscopic microchannel (4,5) after blood flows through microchannel (4,5) fully, carry out the medical analysis judgement.
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Cited By (9)
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| CN104267200A (en) * | 2014-09-17 | 2015-01-07 | 电子科技大学 | Cancer cell detecting micro-fluidic chip based on micro-sized grains on surface of runner and production method |
| CN104293666A (en) * | 2014-09-11 | 2015-01-21 | 大连理工大学 | Micro-fluidic chip device for detecting interaction between two different unicells |
| CN104513787A (en) * | 2015-01-07 | 2015-04-15 | 东北大学 | Integrated micro-fluidic chip and system for capture, culture and administration of single cells |
| CN106434302A (en) * | 2016-09-18 | 2017-02-22 | 华中科技大学 | Portable non-power-source microfluidic cell separation chip |
| CN107262171A (en) * | 2017-07-12 | 2017-10-20 | 华讯方舟科技有限公司 | A kind of blood separation and culture chip and blood separating mechanism |
| CN107702973A (en) * | 2017-09-08 | 2018-02-16 | 深圳市太赫兹科技创新研究院有限公司 | A kind of whole blood blood plasma piece-rate system and method |
| CN109746064A (en) * | 2019-01-28 | 2019-05-14 | 武汉纺织大学 | A kind of gradient magnetic micro-fluidic chip |
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| CN112275337A (en) * | 2020-10-29 | 2021-01-29 | 上海荧辉医疗器械有限公司 | Microfluidic chip and cell screening device and method |
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| CN104293666A (en) * | 2014-09-11 | 2015-01-21 | 大连理工大学 | Micro-fluidic chip device for detecting interaction between two different unicells |
| CN104293666B (en) * | 2014-09-11 | 2016-06-22 | 大连理工大学 | The micro flow control chip device of the interphase interaction that two kinds of differences are unicellular |
| CN104267200A (en) * | 2014-09-17 | 2015-01-07 | 电子科技大学 | Cancer cell detecting micro-fluidic chip based on micro-sized grains on surface of runner and production method |
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| CN104513787A (en) * | 2015-01-07 | 2015-04-15 | 东北大学 | Integrated micro-fluidic chip and system for capture, culture and administration of single cells |
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| CN106434302A (en) * | 2016-09-18 | 2017-02-22 | 华中科技大学 | Portable non-power-source microfluidic cell separation chip |
| CN106434302B (en) * | 2016-09-18 | 2018-03-13 | 华中科技大学 | A kind of micro-current controlled cell separating chips in Portable no-power source |
| CN107262171A (en) * | 2017-07-12 | 2017-10-20 | 华讯方舟科技有限公司 | A kind of blood separation and culture chip and blood separating mechanism |
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| CN107702973A (en) * | 2017-09-08 | 2018-02-16 | 深圳市太赫兹科技创新研究院有限公司 | A kind of whole blood blood plasma piece-rate system and method |
| CN111257205A (en) * | 2018-11-30 | 2020-06-09 | 山东大学 | Cell distribution analysis method |
| CN109746064A (en) * | 2019-01-28 | 2019-05-14 | 武汉纺织大学 | A kind of gradient magnetic micro-fluidic chip |
| CN112275337A (en) * | 2020-10-29 | 2021-01-29 | 上海荧辉医疗器械有限公司 | Microfluidic chip and cell screening device and method |
| CN112275337B (en) * | 2020-10-29 | 2022-03-18 | 上海荧辉医疗器械有限公司 | Microfluidic chip and cell screening device and method |
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Application publication date: 20131113 |