CN104152345A - Microfluidic type single-cell separator - Google Patents
Microfluidic type single-cell separator Download PDFInfo
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- CN104152345A CN104152345A CN201410399750.0A CN201410399750A CN104152345A CN 104152345 A CN104152345 A CN 104152345A CN 201410399750 A CN201410399750 A CN 201410399750A CN 104152345 A CN104152345 A CN 104152345A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M45/00—Means for pre-treatment of biological substances
- C12M45/02—Means for pre-treatment of biological substances by mechanical forces; Stirring; Trituration; Comminuting
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Abstract
The invention discloses a microfluidic single-cell separator, which comprises a base, a movable disk and micro flow channels. The base comprises a motor and a centric crank slider structure connected with the motor, the movable plate is arranged above the base, an inclined plane tilted to one side is above the movable plate, the micro flow channels are arranged on the inclined plane and distributed in the shape of tree forks a branch, and the motor in the base drives the movable plate to conduct translational motion through the centric crank slider structure; the bottom of the movable plate is connected with the centric crank slider structure through a fixed plate; a transparent cover plate is arranged above the micro channels on the inclined plane; the micro channels are made of organic polymer materials; and hollow guide grooves are arranged at the outflow ends of the micro channel. According to the invention, the micro channels are arranged on the inclined plane of the movable plate, and are in the shape of branched tree; the motor inside the base drives the movable plate to conduct translational motion through the centric crank slider structure; the cell suspension in the micro channels flow in the branched channels; and cell suspension in the micro channels isolate single cells under the movement of the movable plate.
Description
Technical field
The present invention relates to a kind of micro-fluidic type single cell separator, belong to biological medicine plant and instrument.
Background technology
The research of current unicellular level more and more comes into one's own, and has been widely used in the fields such as single celled DNA cloning, gene order-checking and the research of unicellular horizontal ubcellular drug delivery system.
Existing unicellular tripping device and method mainly contain: 1, fluorescence Flow Cytometry partition method: fluorescence-activation Flow cytometry can effectively obtain individual cells, but the performance requriements of convection type cell instrument is higher, and instrument is expensive; 2, laser capture microdissection cutting technique: can be from tissue isolated mononuclear cell, but be not suitable for cultivation and the amplification of cell, and need operator extremely expertly to grasp cutting technique; 3, micro-manipulation instrument: adopt micro-manipulation technology to draw from single-cell suspension liquid unicellular, but meticulous operation and valuable instrument are hung back investigator; The defect of above technology makes it to be widely applied.
1998, the micro-processing method of the quick copy moulding that utilizes elasticity macromolecular material PDMS has been invented by the Whitesides seminar of Harvard University, for micro-fluidic chip preparation, the method is called as soft etching technique (soft lithography).With respect to glass material and silicon materials, this technology processing and fabricating is convenient, micro-processing conditions and experiment condition that need not be harsh especially, the template of once making can be repeatedly used, greatly shorten the required time of chip manufacturing, accelerate the speed of research, reduced difficulty and the cost of chip manufacturing.
Polydimethylsiloxane (poly-dimethylsiloxane, PDMS) be that a kind of silicon is basic organic polymer material, be widely used in the fields such as micro-fluidic, biology is micro electronmechanical, can be used as micro sprue system, spackling, lubricant, contact lens etc., there is cost low, use simply, optical clear, and in the ordinary course of things, it is inertia, nontoxic, nonflammable, with there being the features such as good adhesivity between silicon chip.
In view of this, based on this organic polymer material of PDMS, existing unicellular tripping device is improved,, operation high with workout cost is difficult etc., and problem is necessary.
Summary of the invention
The object of the present invention is to provide one can sharp separation unicellular, can in Biohazard Safety Equipment, operate, ensure aseptic technique, part detachable, uses after sterilizing, micro-fluidic type single cell separator simple to operate.
For achieving the above object, the present invention adopts following technical scheme:
A kind of micro-fluidic type single cell separator, comprise base, movable disk and fluid channel, base comprise motor be connected with motor to innermost being handle slide block structure, movable disk is arranged on the top of base, the top of movable disk is the inclined-plane to lopsidedness, fluid channel is set on inclined-plane, the tree fork shape that is distributed as branch of fluid channel, the motor in base is by driving movable disk to carry out translational movement to innermost being handle slide block structure.
Movable disk bottom by retaining plate with innermost being handle slide block structure is connected.
On inclined-plane, the top of fluid channel arranges transparent cover plate.
The material of fluid channel is organic polymer material.
In fluid channel, outflow end has the diversion trench of hollow.
Beneficial effect: fluid channel of the present invention is on the inclined-plane of movable disk, fluid channel is the tree-shaped microchannel of branch, motor in base is by driving movable disk to carry out translational movement to innermost being handle slide block structure, the cell suspending liquid adding in fluid channel moves in the tree-shaped passage of branch, and the cell suspending liquid in fluid channel is unicellular next by isolating in cell suspending liquid under the motion of movable disk.
brief description of the drawings
Fig. 1 is the front view of micro-fluidic type single cell separator;
Fig. 2 is the sectional view that micro-fluidic type single cell separator splits;
Fig. 3 is the fine structure map of fluid channel;
Fig. 4 is that micro-fluidic type single cell separator is isolated the state graph when unicellular.
In figure: 10-base, 101-motor, 102-to innermost being handle slide block structure, 20-movable disk, 201-transparent cover plate, 202-diversion trench, 203-retaining plate, 30-fluid channel, 301-add entrance, 302-spout, 40-unicellular.
Embodiment
In order to make the object, technical solutions and advantages of the present invention clearer, describe the present invention below in conjunction with the drawings and specific embodiments.
As shown in Figures 1 to 4, micro-fluidic type single cell separator of the present invention in order to isolate unicellular 40 from cell suspending liquid.
Micro-fluidic type single cell separator, comprise base 10, movable disk 20 and fluid channel 30, base 10 comprise motor 101 be connected with motor to innermost being handle slide block structure 102, movable disk 20 is arranged on the top of base 10, the top of movable disk 20 is the inclined-plane to lopsidedness, fluid channel 30 is set on inclined-plane, the tree fork shape that is distributed as branch of fluid channel 30, the motor 101 in base 10 is by driving movable disk 20 to carry out translational movement to innermost being handle slide block structure 102.
Innermost being handle slide block structure 102 materials be can be to metal or plastic material, and metal material mainly comprises stainless steel or aluminium, plastic material is mainly polypropylene.In present embodiment, be stainless steel to the material of innermost being handle slide block structure 102.
Described motor 101 and innermost being handle slide block structure 102 is linked, drives movable disk 20 to carry out translational movement, to being connected by retaining plate 203 between innermost being handle slide block structure 102 and movable disk 20.Movable disk 20 materials can be metal or plastic material, and metal material mainly comprises stainless steel or aluminium, and plastic material is mainly polypropylene.In present embodiment, the material of movable disk 20 is polypropylene.
Movable disk 20 appears as ladder structure of right angle from a side, and fluid channel 30 is positioned on the ramp of movable disk 20, and the material of composition fluid channel 30 is organic polymer material, and in present embodiment, the material of composition fluid channel 30 is PDMS.On ramp, there is a transparent cover plate 201, cover and in fluid channel 30, prevent from being polluted in cellular segregation process, be provided with diversion trench 202 at fluid channel 30 ends, convenient reception isolated unicellular 40.
Fluid channel 30 is fixed by the ramp of tackiness agent and movable disk 20.It is tree-shaped that fluid channel 30 is branch, and each section of fluid channel is S type curve tunnel, and the runner circular arc of fluid channel 30 is divided into different size according to cell size, as 100 μ m, and 150 μ m, 200 μ m etc., to reduce Flow Velocity under liquid, facilitate isolated cell.
Cell suspending liquid enters fluid channel 30 by adding entrance 301, cell suspending liquid passes through under the effect of translational movement and gravitional force in the S of fluid channel 30 type curve tunnel, shunt separation, final process diversion trench 202 oozes (each branches end of fluid channel 30 is corresponding one by one with diversion trench) from spout 302, thereby isolates unicellular 40.Ensure aseptic, simple, easy to operate.Micro-fluidic type single cell separator in present embodiment is obtaining after single intact cell, can be used for single celled photoelectric synchronous and detects.
In sum, micro-fluidic type single cell separator of the present invention is by the motor 101 of base 10 inside and the interlock to innermost being handle slide block structure 102, drive the movable disk 20 of the rectangular trapezoidal shape in side to carry out translational movement, be positioned at the fluid channel 30 of the internal passages with the bending of S type on movable disk 20 ramps under the effect of translational movement and gravitional force, cell suspending liquid is separated into unicellular 40.The micro-fluidic type single cell separator of the present invention can operate in Biohazard Safety Equipment, ensure aseptic technique, part detachable, after sterilizing, use, simple to operate, the material of composition fluid channel is New Type Material polydimethylsiloxane (PDMS), cost is low, have good unreactiveness, not damaging cells, is conducive to follow-up cultivation and the detection of cell.
Claims (5)
1. a micro-fluidic type single cell separator, it is characterized in that: comprise base, movable disk and fluid channel, described base comprise motor be connected with motor to innermost being handle slide block structure, described movable disk is arranged on the top of base, the top of movable disk is the inclined-plane to lopsidedness, fluid channel is set on inclined-plane, the tree fork shape that is distributed as branch of described fluid channel, the motor in base is by driving movable disk to carry out translational movement to innermost being handle slide block structure.
2. micro-fluidic type single cell separator according to claim 1, is characterized in that: described movable disk bottom by retaining plate with innermost being handle slide block structure is connected.
3. micro-fluidic type single cell separator according to claim 1, is characterized in that: on described inclined-plane, the top of fluid channel arranges transparent cover plate.
4. micro-fluidic type single cell separator according to claim 1, is characterized in that: the material of described fluid channel is organic polymer material.
5. micro-fluidic type single cell separator according to claim 1, is characterized in that: in described fluid channel, outflow end has the diversion trench of hollow.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201410399750.0A CN104152345B (en) | 2014-08-14 | 2014-08-14 | Micro-fluidic type single cell separator |
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| CN201410399750.0A CN104152345B (en) | 2014-08-14 | 2014-08-14 | Micro-fluidic type single cell separator |
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| CN104152345A true CN104152345A (en) | 2014-11-19 |
| CN104152345B CN104152345B (en) | 2016-08-24 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105950461A (en) * | 2016-07-20 | 2016-09-21 | 江苏瑞明生物科技有限公司 | Single cell nano accurate drug delivery system |
| CN109920482A (en) * | 2019-01-29 | 2019-06-21 | 厦门大学 | A method of analyzing unicellular content |
| CN113322167A (en) * | 2021-04-26 | 2021-08-31 | 上海交通大学 | Micro-fluidic chip |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102472709A (en) * | 2009-07-06 | 2012-05-23 | 索尼公司 | Microfluidic device having onboard tissue or cell sample handling capability |
| CN103131693A (en) * | 2013-02-25 | 2013-06-05 | 宁波美晶医疗技术有限公司 | Separation method of rare cells in blood |
| CN203284394U (en) * | 2013-05-17 | 2013-11-13 | 宁波美晶医疗技术有限公司 | Pendulous device for blood detection biological chips |
| CN203976784U (en) * | 2014-08-14 | 2014-12-03 | 江苏瑞明生物科技有限公司 | micro-fluidic type single cell separator |
-
2014
- 2014-08-14 CN CN201410399750.0A patent/CN104152345B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102472709A (en) * | 2009-07-06 | 2012-05-23 | 索尼公司 | Microfluidic device having onboard tissue or cell sample handling capability |
| CN103131693A (en) * | 2013-02-25 | 2013-06-05 | 宁波美晶医疗技术有限公司 | Separation method of rare cells in blood |
| CN203284394U (en) * | 2013-05-17 | 2013-11-13 | 宁波美晶医疗技术有限公司 | Pendulous device for blood detection biological chips |
| CN203976784U (en) * | 2014-08-14 | 2014-12-03 | 江苏瑞明生物科技有限公司 | micro-fluidic type single cell separator |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105950461A (en) * | 2016-07-20 | 2016-09-21 | 江苏瑞明生物科技有限公司 | Single cell nano accurate drug delivery system |
| CN109920482A (en) * | 2019-01-29 | 2019-06-21 | 厦门大学 | A method of analyzing unicellular content |
| CN113322167A (en) * | 2021-04-26 | 2021-08-31 | 上海交通大学 | Micro-fluidic chip |
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| CN104152345B (en) | 2016-08-24 |
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