CN105907633A - Micro-fluidic chip device used for fine sorting, counting and detection of cell - Google Patents
Micro-fluidic chip device used for fine sorting, counting and detection of cell Download PDFInfo
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- 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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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- 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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/16—Microfluidic devices; Capillary tubes
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
- B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0809—Geometry, shape and general structure rectangular shaped
- B01L2300/0819—Microarrays; Biochips
Abstract
The invention provides a micro-fluidic chip device used for the fine sorting, the counting and the detection of a cell. The micro-fluidic chip device comprises a micro-fluidic chip, a laser counting component and a Raman laser transcribing and detection component, wherein the laser counting component and the Raman laser transcribing and detection component are arranged on one side of the micro-fluidic chip; the micro-fluidic chip comprises an upper-layer auxiliary structure, a middle-layer screening area and a lower-layer sheath flow counting detection area, wherein the upper-layer auxiliary structure covers the top of the middle-layer screening area; the middle-layer screening area covers the top of the lower-layer sheath flow counting detection area. Cells are sorted on the basis of a secondary flow Dejan flow inertial effect and a cylindrical screening structure according to size, and the integrated laser counting component and Raman laser transcribing and detection component can count and detect the sorted cells after cell sorting is finished. The micro-fluidic chip device has a high integration degree and is a multifunctional miniaturized cell sorting, counting and detection device which does not damage the cells and has high cost performance.
Description
Technical field
The present invention relates to micro-fluidic chip and biological medicine research, biochemistry detection technical field, a kind of micro flow control chip device finely sorting for cell, count and detecting based on Secondary Flow Di's grace stream inertia effect.
Background technology
Various cell separation and screening method take on indispensable key player in the fields such as medical diagnosis, clinical treatment, cytobiology.Currently used relatively broad cell sorting method is membrane filter method and flow cytometry, and the disadvantage of membrane filter method is that fenestra is easily blocked by cell, causes separation efficiency reduction, sample loss and pollution etc..The major defect of flow cytometry (fluorescence amplifying cell separator) is that instrument is relatively big, need that sample dose is big, operation complexity, relies only on central laboratory, professional manipulates, and operating process affects cytoactive.The magnetic activated cell separating method developed by conventional flow cell analysis art and come, has flux height, high specificity and separating purity advantages of higher.However it is necessary that the antibody modification that target cell is carried out uniqueness, add the complexity of cell sorting process, extend experimental period, most importantly destroy the initial condition of cell, thus constrain promoting the use of of this technology.Additionally, above-mentioned cell isolation method can not be integrated with other analysis and detection device or combine, it is impossible to being applied to build increasingly complex detection and analysis system, the most existing cell sorting method cannot meet the demand of cell detection with rapid changepl. never-ending changes and improvements.
Micro-fluidic chip is a kind of micro devices that can operate trace level sample, is integrated with many tiny microchannels, in order to fluid passes through and operates on it on micro-fluidic chip.
Micro-fluidic chip arranges bending channel, when fluid flows through bending channel, in addition to travelling forward along bending channel, the cross section be perpendicular to flow direction can be formed a pair reverse eddy current, they at the middle part of bending channel to tube wall medial motion, and in top and bottom to tube wall movement outside, this flowing is referred to as Secondary Flow Di's grace stream.Under the effect of Di's grace stream inertia effect, the cell in fluid is sized in bending channel cross section, and travelling forward along bending channel forms respective flow route, finally in the sorting passage that exit is different according to the difference entrance of horizontal level.
The advantage utilizing Di's grace stream inertia effect to carry out cell sorting is: utilize high flow rate to realize Di's grace stream inertia effect, and in microchannel, microgranule or cell to certain size carry out flowing arrangement, substantially increase the flux processing sample；Without applying any external force such as electromagnetic force, mechanical force etc. during sorting, without compromising on cell viability；Chip apparatus simple in construction, volume is little, it is not necessary to mechanically or electrically subassembly, it is easy to be combined with more existing chip lab system integrations as functional module.By associated advantages, the cell sorting micro-fluidic chip of Di's grace stream inertia effect is utilized to be used widely in the field such as bioanalysis and clinical medicine.
Finding by consulting domestic and international pertinent literature and autonomic nerve way, the most various cell sorting micro-fluidic chips based on Di's grace stream effect have the higher efficiency of separation, typically can reach more than 80%；But only under the effect of Di's grace stream inertia effect, all the time the cell having other sizes of minority is mingled in the cell flow route of a certain single size, enter in the sorting passage specified, reduce final sorting index, autonomic nerve way have also discovered this phenomenon.
Summary of the invention
It is an object of the invention to provide a kind of micro flow control chip device finely sorting for cell, count and detecting, this device uses micro-fluidic chip based on Secondary Flow Di's grace stream inertia effect and cylindricality screening structure to sort cell by size, and integrated laser counter block, raman laser adopt spectrum and detection part, after cell sorting completes, the cell sorted can be counted and detects.
The technical scheme is that
A kind of micro flow control chip device finely sorting for cell, count and detecting, the laser counter block including micro-fluidic chip and being arranged on described micro-fluidic chip side and raman laser adopt spectrum and detection part, described micro-fluidic chip includes upper strata supplementary structure, screening area, middle level and lower floor's sheath flow accounting detection zone, described upper strata supplementary structure covers on screening area, middle level, and screening area, described middle level covers on lower floor's sheath flow accounting detection zone；
Described upper strata supplementary structure is provided with treats sorted sample liquid top-level entries, clamping stream top-level entries, the outlet of remaining sample liquid upper strata and the outlet of sheath flow liquid upper strata, screening area, described middle level is provided with treats that sorted sample liquid middle level entrance, the outlet of monotonous helical shape bending micro, sorted sample liquid middle level, the outlet of remaining sample liquid middle level, clamping stream middle level are crossed ferry and sheath flow liquid middle level and crossed ferry, and described lower floor sheath flow accounting detection zone is provided with sorted sample liquid lower floor entrance, clamping and flows down a layer entrance, microchannel and the outlet of sheath flow liquid lower floor；
Described treat sorted sample liquid top-level entries and treat that sorted sample liquid middle level entrance connects, the outlet of the described liquid of sorted sample middle level connects with sorted sample liquid lower floor entrance, the outlet of described remaining sample liquid middle level and remaining sample liquid upper strata outlet, described clamping stream top-level entries is crossed ferry by clamping stream middle level and is flowed down a layer entrance with clamping and connect, and described sheath flow liquid lower floor exports crosses ferry and sheath flow liquid upper strata outlet by sheath flow liquid middle level；
Described monotonous helical shape bending micro has several spiral coils, each spiral coil is equipped with cylindricality screening structure, described cylindricality screening structure is made up of the cylinder that several rows are evenly distributed, and the cylindricality screening structure in each spiral coil from inside to outside, the distance between its cylinder is sequentially reduced；The inner side of described monotonous helical shape bending micro or outside are provided with side passage, and one end of described side passage connects with monotonous helical shape bending micro, the other end and sorted sample liquid middle level outlet；
Described lower floor sheath flow accounting detection division is divided into focal zone, counting region and adopts spectrum detection region, the described liquid of sorted sample and clamping stream pool sheath stream at focal zone, described sheath stream, for being focused the same size cell sorted, makes same size cells in alignment detect region by counting region with adopting spectrum one by one.
The described micro flow control chip device finely sorting for cell, count and detecting, described laser counter block includes laser instrument, signal receiver and data handling system, it is incident that the laser beam of described laser instrument output is perpendicular to counting region, and the outfan of described signal receiver is connected with the input of data handling system.
The described micro flow control chip device finely sorting for cell, count and detecting, described raman laser is adopted spectrum and is included Raman laser light source, detector and Raman signal processing system with detection part, the laser beam of described Raman laser light source output is perpendicular to adopt spectrum detection region incidence, and the outfan of described detector is connected with the input of Raman signal processing system.
The described micro flow control chip device finely sorting for cell, count and detecting, the microchannel of described focal zone is class cross, is made up of with being arranged on both sides, main channel and angle two wing passages less than 90 degree main channel；One end of described main channel connects with sorted sample liquid lower floor entrance, and other end one end with two wing passages respectively connects, and the other end of said two wing passage connects with clamping inflow entrance respectively.
The described micro flow control chip device finely sorting for cell, count and detecting, described counting region and the microchannel adopting spectrum detection region are linear.
The described micro flow control chip device finely sorting for cell, count and detecting, described upper strata supplementary structure, screening area, middle level and lower floor's sheath flow accounting detection zone are rectangle, wherein, screening area, described middle level and lower floor's sheath flow accounting detection zone are constituted by one block of overall PDMS plate.
The invention have the benefit that
As shown from the above technical solution, the invention provides a kind of integrated level higher, not damaging cells, high performance-price ratio, multi-functional, the cell sorting detection instrument of miniaturization, micro-fluidic chip in the present invention is without applying any external force such as electromagnetic force, mechanical force etc., cell can be sorted by Di's grace stream inertia effect by size that only utilize the monotonous helical shape bending micro characteristic of self to be formed, and by increasing cylindricality screening structure and side passage in each spiral coil of monotonous helical shape bending micro, further increase the efficiency carrying out cell sorting based on Di's grace stream inertia effect, thus improve accuracy and the reliability of micro-fluidic chip sorting cells.
Multiple analysis detection means is combined by the present invention, utilize sheath stream that cell is focused, utilize laser that the cell after sorting, focusing is counted, utilize Raman spectrum that cell is sampled, it is a kind of complex multi-functional, highly integrated micro-fluidic chip detection and analysis system, have quickly, high flux, high efficiency, the feature such as multi-functional, integrated and miniaturization, be expected to be used widely in the field such as bioanalysis and clinical medicine.
Accompanying drawing explanation
Fig. 1 is the overall structure schematic diagram of the micro-fluidic chip of the present invention；
Fig. 2 is the structural representation of the monotonous helical shape bending micro of the micro-fluidic chip of the present invention；
Fig. 3 is cylindricality screening structure and the distribution schematic diagram of side passage of the micro-fluidic chip of the present invention；
Fig. 4 is that the micro-fluidic chip of the present invention utilizes bending micro to form the principle schematic of Di's grace stream inertia effect；
Fig. 5 is that the micro-fluidic chip of the present invention utilizes cylindricality screening structure to remove the schematic diagram of other size cells of minority in single size cell flow route；
Fig. 6 is that the micro-fluidic chip of the present invention utilizes sheath stream that cell focuses on the principle schematic being arranged in rows；
Fig. 7 is the laser counter block structural representation of the present invention；
Fig. 8 is that the raman laser of the present invention adopts spectrum and detection part structural representation.
Detailed description of the invention
The present invention is further illustrated below in conjunction with the accompanying drawings with specific embodiment.
As shown in Figure 1 and Figure 2, a kind of micro flow control chip device finely sorting for cell, count and detecting, adopts spectrum and detection part 5 including the micro-fluidic chip being made up of upper strata supplementary structure 1, screening area, middle level 2 and lower floor's sheath flow accounting detection zone 3 and the laser counter block 4 being arranged on micro-fluidic chip side, raman laser.Upper strata supplementary structure 1 covers on screening area, middle level 2, and screening area, middle level 2 covers on lower floor's sheath flow accounting detection zone 3.
Upper strata supplementary structure 1 is one piece of rectangular glass cover-plate, and it has a lot of micropore, it is simple to connect micro-pipe.Upper strata supplementary structure 1 plays support, seals screening area, middle level 2 and the effect of lower floor's sheath flow accounting detection zone 3, offers simultaneously and need sorted sample liquid top-level entries 11, clamping stream top-level entries 12, remaining sample liquid upper strata outlet 13 and sheath flow liquid upper strata outlet 14 on upper strata supplementary structure 1.Wherein, treating that sorted sample liquid top-level entries 11 is the interface that the sample liquid comprising and treating sorting cells flows into from the external world, clamping stream top-level entries 12 is the interface that clamping stream flows into from the external world；Remaining sample liquid upper strata outlet 13 is the interface gone out through the remaining sample liquid stream sorted；The interface that sheath flow liquid upper strata outlet 14 is flowed out for sheath flow liquid.
Screening area, middle level 2 is rectangle, and is made up of one block of overall PDMS plate, it has a lot of MCA, is mainly responsible for realizing treating in microchannel that sorted sample liquid stream leads to, the cell treated in sorted sample liquid carries out the function that sorts.Screening area, middle level 2 includes some treating that sorted sample liquid middle level entrance 21, monotonous helical shape bending micro 22, the some liquid of sorted sample middle levels outlet 23, remaining sample liquid middle level outlet 24, clamping stream middle level are crossed ferry 25 and sheath flow liquid middle level and crossed ferry 26.Wherein, treat sorted sample liquid middle level entrance 21 and treat that sorted sample liquid top-level entries 11 connects, for treating that sorted sample liquid stream enters the interface of screening area, middle level 2；Monotonous helical shape bending micro 22 has certain rotating cycle, and the species number of cell that the concrete number of turns carries out sorting as required is configured, and utilizes monotonous helical shape bending micro 22 can form Secondary Flow Di's grace stream inertia effect and sorts cell by size；Sorted sample liquid middle level outlet 23 is the interface that the sample liquid comprising and sorting single size cell flows out screening area, middle level 2；Remaining sample liquid middle level outlet 24 connects with remaining sample liquid upper strata outlet 13, for going out the interface of screening area, middle level 2 through the remaining sample liquid stream of sorting；Clamping stream middle level is crossed ferry 25 and is connected with clamping stream top-level entries 12, flows into the changeover joint of lower floor's sheath flow accounting detection zone 3 from upper strata supplementary structure 1 for clamping stream；Sheath flow liquid middle level is crossed ferry 26 and is connected with sheath flow liquid upper strata outlet 14, flows into the changeover joint of upper strata supplementary structure 1 from lower floor's sheath flow accounting detection zone 3 for sheath flow liquid.
Lower floor's sheath flow accounting detection zone 3 is rectangle, it is made up of one block of overall PDMS plate equally, there are some MCAs on it, are mainly responsible for realizing the function that sorted sample liquid circulates in microchannel, the cell in sorted sample liquid is focused, counts and is detected.Some sorted sample liquid lower floor entrance 31 and some sheath flow liquid lower floors outlet 32 is offered on lower floor's sheath flow accounting detection zone 3.Wherein, sorted sample liquid lower floor entrance 31 connects with the outlet of sorted sample liquid middle level 23, flows into the interface of lower floor's sheath flow accounting detection zone from screening area, middle level 2 for comprising the sample liquid sorting single size cell；Sheath flow liquid lower floor outlet 32 is crossed ferry 26 with sheath flow liquid middle level and is connected, the interface flowed out from lower floor's sheath flow accounting detection zone 3 for sheath flow liquid.
As shown in Figure 3, the a certain region of monotonous helical shape bending micro 22 is provided with cylindricality screening structure, including the first cylindricality screening structure the 27, second cylindricality screening structure the 28, the 3rd cylindricality screening structure 29 and the 4th cylindricality screening structure 210, cylinder by several rows marshalling is constituted, from inside to outside, the distance being arranged between the cylinder constituting cylindricality screening structure in the different spiral coil of monotonous helical shape bending micro 22 is gradually reduced, the main cell being responsible for removing other sizes of minority in the cell flow route of single size.Wherein, first cylindricality screening structure 27 is arranged in first spiral coil of monotonous helical shape bending micro 22, the distance constituted between the cylinder of the first cylindricality screening structure 27 is maximum, second cylindricality screening structure the 28, the 3rd cylindricality screening structure 29 and the 4th cylindricality screening structure 210 are successively set in second to the 4th spiral coil of monotonous helical shape bending micro 22, and the distance constituted between the second cylindricality screening structure the 28, the 3rd cylindricality screening structure 29 and the cylinder of the 4th cylindricality screening structure 210 is sequentially reduced.The inner side (or outside) of monotonous helical shape bending micro 22 is provided with side passage 211, mainly it is responsible for will not pass through cylindricality screening structure, and is exported to lower floor sheath flow accounting detection zone 3 sideways against the cell acted on via the outlet of sorted sample liquid middle level 23 by Di Enli.
As shown in Figure 4, when when sorted sample liquid stream is through monotonous helical shape bending micro 22, in addition to travelling forward along runner, the cross section of monotonous helical shape bending micro 22 can be formed a pair relative Di's grace stream 213 of flow direction, they in the middle part of runner to tube wall medial motion, and in top and bottom to tube wall movement outside.The cross section of monotonous helical shape bending micro 22 is perpendicular to treat sorted sample liquid flow direction 212.
Under the effect of Di's grace stream inertia effect, treating that the cell in sorted sample liquid is sized in the cross section of monotonous helical shape bending micro 22, travelling forward along monotonous helical shape bending micro 22 forms respective flow route.When various sizes of cell moves forward to, along respective flow route, the region being provided with cylindricality screening structure, the cell that size is less than the spacing of the cylinder of cylindricality screening structure is able to screen structure by cylindricality, enter in the next spiral coil of monotonous helical shape bending micro 22, the cell that size is bigger than the spacing of the cylinder of cylindricality screening structure then can not screen structure by cylindricality, and Di's grace power 214 on the cross section of monotonous helical shape bending micro 22 sideways against effect under, enter in the side passage 211 being arranged on monotonous helical shape bending micro 22 side, prevent cell from reuniting near cylindricality screening structure, form blocking.
Due to from inside to outside, the distance constituted on each spiral coil of monotonous helical shape bending micro 22 between the cylinder of cylindricality screening structure is gradually reduced, first the cell that size is maximum is isolated at first spiral coil, first isolate, at second spiral coil, the cell that size is slightly smaller, sub-elect, in rear several spiral coils, the cell that size is gradually reduced successively.The cylindricality screening structure arranged in each spiral coil of monotonous helical shape bending micro 22 has effectively filtered the cell of other sizes of minority in the cell flow route being mingled in a certain single size, therefore further increase the efficiency carrying out cell sorting based on Di's grace stream effect, improve levels of precision and the reliability of micro-fluidic chip sorting cells.
As shown in Figure 5, when large scale cell 61, medium size cell 62, small size cell 63 and minimum dimension cell 64 along treat sorted sample liquid flow direction 212 move forward to be arranged in first spiral coil of monotonous helical shape bending micro 22 first cylindricality screening structure 27 before, constitute first cylindricality screening structure 27 cylinder between distance be maximum.Therefore, large scale cell 61 will be unable to by the first cylindricality screening structure 27, and less medium size cell 62, small size cell 63 and the minimum dimension cell 64 of size will be through the first cylindricality screening structure 27 along treating in the next spiral coil that sorted sample liquid flow direction 212 enters into monotonous helical shape bending micro 22.The large scale cell 61 in the next spiral coil of monotonous helical shape bending micro 22 cannot can not be entered into through the first cylindricality screening structure 27, Di's grace power 214 on the cross section of monotonous helical shape bending micro 22 sideways against effect under, enter in the side passage 211 being arranged on monotonous helical shape bending micro 22 side, and enter lower floor's sheath flow accounting detection zone 3 by corresponding sorted sample liquid middle level outlet 23.
As shown in Figure 6, lower floor's sheath flow accounting detection zone 3 is divided into focal zone 34, counting region 35 and adopts spectrum detection region 36.In focal zone 34, microchannel is shaped as Y-shaped cross, when comprising the liquid of sorted sample of the same size cell 65 after having classifyd by size when sorted sample liquid lower floor entrance 31 flows into the main channel of focal zone 34, cross ferry 25 from clamping stream top-level entries 12 via clamping stream middle level and flow down layer entrance 33 input clamping stream to clamping, clamping stream flows into main channel from two wing passages of focal zone 34, converges with sorted sample liquid and forms sheath stream.Sheath stream refers to that fluid, under lateral compression effect effect, forms the compression flow form being similar to sheath shape.By sheath stream, the same size cell 65 sorted is focused, makes same size cell 65 be arranged in rows one by one and detect region 36 by counting region 35 with adopting spectrum.Counting region 35 and adopt microchannel in spectrum detection region 36 be shaped as orthoscopic, predominantly sorting, focus on after the counting of cell and adopt spectrum detection working place be provided.
As it is shown in fig. 7, laser counter block 4 includes laser instrument 41, signal receiver 42 and data handling system 43, it is incident that the laser beam of laser instrument 41 output is perpendicular to counting region 35.Laser instrument 41 continuously sends out laser signal, when same size cell 65 is by laser beam, light can produce refraction and reflection, collect refraction and the signal of reflection by signal receiver 42, and in data handling system 43, the form with a peak value carries out record.Often by one with size cell 65, a peak value, the total degree that finally record peak value occurs will be produced, i.e. obtain the number of same size cell 65.
As shown in Figure 8, raman laser is adopted spectrum and is included Raman laser light source 51, detector 52 and Raman signal processing system 53 with detection part 5, and it is incident that the laser beam that Raman laser light source 51 exports is perpendicular to adopt spectrum detection region 36.Raman laser light source 51 sends laser signal, when same size cell 65 is by raman laser bundle, recording same size cell 65 by detector 52 makes incident ray that the Raman signal of scattering to occur, by Raman signal processing system 53, Raman signal is processed, complete adopting spectrum and detecting same size cell 65.
The above embodiment is only to be described the preferred embodiment of the present invention; not the scope of the present invention is defined; on the premise of designing spirit without departing from the present invention; various deformation that technical scheme is made by those of ordinary skill in the art and improvement, all should fall in the protection domain that claims of the present invention determines.
Claims (6)
1. the micro flow control chip device finely sorting for cell, count and detecting, it is characterized in that: include micro-fluidic chip and be arranged on the laser counter block of described micro-fluidic chip side and raman laser adopts spectrum and detection part, described micro-fluidic chip includes upper strata supplementary structure, screening area, middle level and lower floor's sheath flow accounting detection zone, described upper strata supplementary structure covers on screening area, middle level, and screening area, described middle level covers on lower floor's sheath flow accounting detection zone；
Described upper strata supplementary structure is provided with treats sorted sample liquid top-level entries, clamping stream top-level entries, the outlet of remaining sample liquid upper strata and the outlet of sheath flow liquid upper strata, screening area, described middle level is provided with treats that sorted sample liquid middle level entrance, the outlet of monotonous helical shape bending micro, sorted sample liquid middle level, the outlet of remaining sample liquid middle level, clamping stream middle level are crossed ferry and sheath flow liquid middle level and crossed ferry, and described lower floor sheath flow accounting detection zone is provided with sorted sample liquid lower floor entrance, clamping and flows down a layer entrance, microchannel and the outlet of sheath flow liquid lower floor；
Described treat sorted sample liquid top-level entries and treat that sorted sample liquid middle level entrance connects, the outlet of the described liquid of sorted sample middle level connects with sorted sample liquid lower floor entrance, the outlet of described remaining sample liquid middle level and remaining sample liquid upper strata outlet, described clamping stream top-level entries is crossed ferry by clamping stream middle level and is flowed down a layer entrance with clamping and connect, and described sheath flow liquid lower floor exports crosses ferry and sheath flow liquid upper strata outlet by sheath flow liquid middle level；
Described monotonous helical shape bending micro has several spiral coils, each spiral coil is equipped with cylindricality screening structure, described cylindricality screening structure is made up of the cylinder that several rows are evenly distributed, and the cylindricality screening structure in each spiral coil from inside to outside, the distance between its cylinder is sequentially reduced；The inner side of described monotonous helical shape bending micro or outside are provided with side passage, and one end of described side passage connects with monotonous helical shape bending micro, the other end and sorted sample liquid middle level outlet；
Described lower floor sheath flow accounting detection division is divided into focal zone, counting region and adopts spectrum detection region, the described liquid of sorted sample and clamping stream pool sheath stream at focal zone, described sheath stream, for being focused the same size cell sorted, makes same size cells in alignment detect region by counting region with adopting spectrum one by one.
The micro flow control chip device finely sorting for cell, count and detecting the most according to claim 1, it is characterized in that: described laser counter block includes laser instrument, signal receiver and data handling system, it is incident that the laser beam of described laser instrument output is perpendicular to counting region, and the outfan of described signal receiver is connected with the input of data handling system.
The micro flow control chip device finely sorting for cell, count and detecting the most according to claim 1, it is characterized in that: described raman laser is adopted spectrum and included Raman laser light source, detector and Raman signal processing system with detection part, the laser beam of described Raman laser light source output is perpendicular to adopt spectrum detection region incidence, and the outfan of described detector is connected with the input of Raman signal processing system.
The micro flow control chip device finely sorting for cell, count and detecting the most according to claim 1, it is characterized in that: the microchannel of described focal zone is class cross, be made up of with being arranged on both sides, main channel and angle two wing passages less than 90 degree main channel；One end of described main channel connects with sorted sample liquid lower floor entrance, and other end one end with two wing passages respectively connects, and the other end of said two wing passage connects with clamping inflow entrance respectively.
The micro flow control chip device finely sorting for cell, count and detecting the most according to claim 1, it is characterised in that: described counting region and the microchannel adopting spectrum detection region are linear.
The micro flow control chip device finely sorting for cell, count and detecting the most according to claim 1, it is characterized in that: described upper strata supplementary structure, screening area, middle level and lower floor's sheath flow accounting detection zone are rectangle, wherein, screening area, described middle level and lower floor's sheath flow accounting detection zone are constituted by one block of overall PDMS plate.
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| CN106582899A (en) * | 2016-11-01 | 2017-04-26 | 深圳市星国华先进装备科技有限公司 | Liquid droplet microfluidic device |
| CN107058060A (en) * | 2017-04-10 | 2017-08-18 | 东南大学 | A kind of tumour cell high flux sorting enrichment micro-fluidic chip |
| CN107159326A (en) * | 2017-05-09 | 2017-09-15 | 西北工业大学 | Inertia based on enhanced Secondary Flow focuses on chip |
| CN110186836A (en) * | 2019-06-21 | 2019-08-30 | 山东师范大学 | The optofluidic flow cytometer of circulating tumor cell separation analysis and Classification Count |
| CN110639628A (en) * | 2019-09-12 | 2020-01-03 | 山东大学 | Micro-fluidic chip and method for cell sorting and positioning |
| CN112708536A (en) * | 2021-01-25 | 2021-04-27 | 中国计量大学 | Cell sorting device |
| CN114196725A (en) * | 2021-12-03 | 2022-03-18 | 清华大学 | Method for identifying microorganism and system thereof |
| CN114260037A (en) * | 2021-12-27 | 2022-04-01 | 杭州电子科技大学 | 3D stacked multistage inertial microfluidic sorting chip and preparation method thereof |
| CN115055213A (en) * | 2022-04-14 | 2022-09-16 | 四川轻化工大学 | Filtering type microfluidic bacterial chip based on SERS detection technology and use method |
| WO2023093404A1 (en) * | 2021-11-23 | 2023-06-01 | 中国科学院青岛生物能源与过程研究所 | Microparticle self-entrainment type microfluidic chip and manufacturing method therefor, and microparticle self-dispersion method |
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