CN112774742B - Blood cell filters washing unit - Google Patents
Blood cell filters washing unit Download PDFInfo
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- CN112774742B CN112774742B CN201911083013.9A CN201911083013A CN112774742B CN 112774742 B CN112774742 B CN 112774742B CN 201911083013 A CN201911083013 A CN 201911083013A CN 112774742 B CN112774742 B CN 112774742B
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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
- B01L3/502753—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 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
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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
- B01L3/502761—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 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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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
- 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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- 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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/14—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes
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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
- C12M39/00—Means for cleaning the apparatus or avoiding unwanted deposits of microorganisms
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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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- 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
- B01L2200/06—Fluid handling related problems
- B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
Abstract
The invention provides a blood cell filtering and washing device which comprises a chip, a filter membrane, a filtering clamping component and a washing clamping component, wherein the chip sequentially comprises a main sealing layer, a first chip and a second chip, the filter membrane is positioned between the first chip and the second chip, the first chip and the second chip comprise hollow structures for exposing the filter membrane, the filtering clamping component comprises a first filtering clamping layer in clamping contact with the sealing layer and a second filtering clamping layer in clamping contact with the second chip, the washing clamping component comprises a first washing clamping layer in clamping contact with the sealing layer and a second washing clamping layer in clamping contact with the second chip, and the filter membrane is irreversibly bonded with the first chip and the second chip. According to the invention, through the flow channel structure, the integration effect of the structure and the subsequent units is effectively improved, and the cell loss is reduced.
Description
Technical Field
The invention provides a cell separation device, in particular to a blood cell filtering and flushing device.
Background
Circulating Tumor Cells (CTCs) and Circulating Tumor Microemboli (CTMs) present in peripheral blood are considered to be important causes and markers of tumor metastasis and recurrence. Its presence both represents the invasive capacity of the primary tumor and predicts the possibility of metastatic formation at distant sites. Therefore, accurate detection of the number of CTCs and CTMs in a unit volume of blood can provide an important basis for early screening of cancer; providing dynamic monitoring and prognosis basis for the disease development of the tumor patient; meanwhile, a research window can be provided for the pathogenic mechanism and the drug resistance mechanism of the tumor cells. The number of CTCs and CTMs per volume is extremely rare relative to other normal blood cells present in peripheral blood, with a ratio of about 1: 109Therefore, accurate screening, enrichment and release of target cells are the premise for reliable detection. In other words, the capture rate and release rate of target cells are important indicators affecting CTCs and CTMs.
Currently, membrane filtration technology (ISET) based on morphological enrichment is one of the major methods for detecting CTCs and CTMs. Has the advantages of simple operation method, lower cost, high biological activity of the enriched CTC and CTM, and the like. However, the diameter of CTC cells is similar to that of some blood cells, and the use of the ISET technology only results in a reduction in enrichment purity, so that the ISET technology requires transfer of CTCs and CTMs enriched by the microporous filter membrane from the filter membrane to other operation units to improve the enrichment index. Because the processing unit of the microfluidic technology has the advantages of less reagent consumption, short reaction and analysis time, low cost, easy integration and the like, the processing unit is often used as a subsequent processing unit. However, this technique also has some technical disadvantages, such as that the filtered cells are not easily separated from the microporous filtration membrane; the input of the initial sample is incompatible with the processing capacity of the microfluidic cell; poor module compatibility results in separation of the steps of filtration and subsequent processing and loss of cells.
Disclosure of Invention
Aiming at the defects that the cells are not easy to separate from the microporous filter membrane after filtration and the cells are lost during separation in the prior art, the invention provides a blood cell filtration and flushing device.
The invention provides a blood cell filtering and separating device, which comprises a chip, a filter membrane, a filtering clamping part and a flushing clamping part, wherein the chip sequentially comprises a sealing layer, a first chip and a second chip; the filtering clamping component comprises a first filtering clamping layer in clamping contact with the sealing layer and a second filtering clamping layer in clamping contact with the second chip; the first filter clamping layer comprises a sample inlet corresponding to the filter membrane; the second filtering and clamping layer comprises a filtrate outlet corresponding to the filter membrane; the flushing clamping component comprises a first flushing clamping layer in clamping contact with the sealing layer and a second flushing clamping layer in clamping contact with the second chip; the first clamping layer that washes includes the lateral flushing liquid entry that corresponds to filter membrane one side position department, and the first clamping device that washes includes the flush fluid export that corresponds to filter membrane opposite side position department, the second wash the clamping layer including corresponding to perpendicular flush fluid entry and the gas vent of filter membrane top department, its improvement lies in: the first flushing clamping layer comprises a flushing liquid flow channel for communicating the lateral flushing liquid inlet and the flushing liquid outlet; the filter membrane is irreversibly bonded with the first chip and the second chip.
Preferably, the thickness of the second chip and the second chip is 0.6mm-3 mm; the effective area of the sample inlet and the sample outlet corresponding to the filter membrane is less than 1cm2(ii) a The diameters of the lateral flushing liquid inlet, the vertical flushing liquid inlet and the flushing liquid outlet are 1.5-2.5 mm; the diameter of the exhaust port is 0.2 mm-1 mm.
Preferably, the wash fluid flow path is in a different plane to the filter membrane. First filtration centre gripping layer is provided with the valve region including closed valve on the flush fluid runner, and the diameter in valve region is 1.1~2 times of flush fluid runner width, and the degree of depth direction be with closed valve curved surface complex arc curved surface. The flushing liquid flow passage comprises a flushing liquid inlet flow passage connected with the lateral flushing liquid inlet and a flushing liquid outlet flow passage connected with the flushing liquid outlet.
The washing liquid inlet flow passage is 2 or more, the washing liquid outlet flow passage is one, and the width ratio of the washing liquid outlet flow passage to the washing liquid inlet flow passage is the inverse ratio of the number of the washing liquid outlet flow passages to the number of the washing liquid inlet flow passages.
Preferably, the irreversible bonding comprises thermocompression bonding. The first chip and the second chip are PMMA, and the ultrasonic bonding treatment comprises the steps of placing a filter membrane material between the first chip and the second chip, fixing the filter membrane material, placing the filter membrane material into an ethanol or acetone solvent, and carrying out ultrasonic treatment on the filter membrane material at 100W-360W for no more than 3 minutes at the temperature lower than 45 ℃; or using 256nm, 0.1J/cm before thermocompression bonding2-10J/cm2The first chip and the second chip are irradiated with the ultraviolet light.
The technical scheme provided by the invention has the following beneficial effects: the technical scheme provided by the invention integrates the microporous filter membrane into the microfluidic chip, and the defects that the release rate of cells on the microporous filter membrane is low and the dosage of an initial sample is not matched with the bearing capacity of a subsequent processing unit are overcome by the different flow channels arranged in the way of the vertical sample inlet and the lateral washing buffer inlet; the problems of separation of processing steps, cell loss and the like caused by poor compatibility of an upstream module and a downstream module are solved.
Drawings
FIG. 1 Split front view of a die before bonding
FIG. 2: chip integral structure diagram after bonding
FIG. 3: filter clamping structure diagram
FIG. 4: flushing clamping structure diagram
FIG. 5: schematic diagram of chip-matched filtering and clamping component
FIG. 6: chip cooperation washing clamping part use schematic diagram
FIG. 7: overlook structure diagram of first flushing clamping layer
FIG. 8: another overlook structure chart of the first washing clamping layer;
1. the chip comprises a chip, 2, a filter membrane, 3, a filtering clamping component, 4, a flushing clamping component, 11, a first chip, 12, a second chip, 13, a sealing layer, 111, a first chip sample filtering inlet, 112, a first chip chamfer, 121, a second chip filtrate outlet, 31, a first filtering clamping layer, 32, a second filtering clamping layer, 41, a first flushing clamping layer, 42, a second flushing clamping layer, 411, a first flushing clamping layer side flushing liquid inlet, 412, a first flushing clamping layer flushing liquid outlet, 413, a first flushing clamping layer flushing liquid inlet flow channel; 414 a first rinsing clamping layer rinsing chamber; 415. a first flushing clamping layer flushing fluid outlet flow channel; 416. a first flushing clamping layer valve area, 421, a second flushing clamping layer flushing liquid vertical inlet; 422. a second purge clamping layer exhaust.
Detailed Description
The invention provides a hemocyte filtration and flushing device which comprises a chip 1, a filter membrane 2, a filtration clamping part 3 and a flushing clamping part 4; as shown in fig. 1, the chip 1 includes a first chip 11, a second chip 12, and a sealing layer 13; the filter membrane 2 is positioned between the first chip 11 and the second chip 12, and the hollow structure between the first chip 11 and the second chip 12 is exposed from the filter membrane 2; as shown in fig. 3, the filter holding member 3 includes a first filter holding layer 31 in holding contact with the sealing layer 13, and a second filter holding layer 32 in holding contact with the second chip 12, the first filter holding layer 31 corresponding to the sample inlet 111 of the filter membrane of the first chip 11, and the second filter holding layer 32 corresponding to the filtrate outlet 121 of the filter membrane 2; as shown in fig. 4, the rinsing nip member 4 includes a first rinsing nip layer 41 in direct contact with the sealing layer 13 and a second rinsing nip layer 42 in nip contact with the second chip 12, the first rinsing nip layer 41 including a rinsing liquid side inlet 411 and a rinsing liquid outlet 412; the second rinsing clamping layer 42 comprises a vertical rinsing liquid inlet 421 corresponding to the position above the filter membrane and an exhaust 422 for keeping pressure balance, and the connection surface of the first chip, the filter membrane and the outlet side is provided with a chamfer 112 to prevent the formation of a liquid flow dead zone.
The flushing liquid flow channel of the first flushing clamping layer is directly contacted with the sealing layer 13 of the chip 1 and is positioned on a different plane from the filter membrane 2; the rinsing liquid flow passage of the first rinsing nip comprises a rinsing liquid inlet flow passage 413 connected to the rinsing liquid side inlet 411 and a rinsing liquid outlet flow passage 415 connected to the rinsing liquid outlet 412.
For the assurance flush fluid smooth flow, the width ratio of the flush fluid outlet flow channel of first washing centre gripping layer and the flush fluid inlet flow channel of first washing centre gripping layer is the inverse ratio of the number of the flush fluid outlet flow channel of first washing centre gripping layer and the flush fluid inlet flow channel of first washing centre gripping layer.
The encapsulation between the first chip 11 and the second chip 12 and between the second chip 12 and the filter membrane 2 are irreversible bonding, and the reversible bonding is between the chip and the filter clamping device.
The microporous filter membrane material of the invention is parylene, Polycarbonate (PC) or polyethylene terephthalate (TETP). The material of the chip is PMMA (polymethyl siloxane, PDMS or polydimethylsiloxane).
The irreversible bonding between the chip and the microporous filter membrane material comprises:
1. placing microporous filter membrane material between chips, fixing with clean clamp, placing into ethanol or acetone solvent, and performing ultrasonic treatment at a temperature below 45 deg.C and 100W-360W for less than 3 min;
2. bonding pretreatment: at a wavelength of 256nm and an intensity of 0.1J/cm2-10J/cm2And irradiating under ultraviolet light to perform thermocompression bonding.
In one embodiment of the invention, the microporous filter membrane material on the chip and the bottom layer channel are not in the same plane, and the first chip is sealed by the sealing layer 13 to wash the microchannel of the clamping layer 41; meanwhile, in order to ensure that the sealing layers 13 and 41 are tightly connected and cannot cause liquid leakage, the sealing layer 13 is made of PDMS, PVC or PET flexible film or PMMA or PC hard material subjected to sealing surface treatment.
In one embodiment of the invention, the manufacturing process of the chip comprises the steps of taking PMMA as a chip material and taking PDMS as a flexible film material with the thickness of 150-300 mu m, wherein the mass ratio of the flexible film material to a curing agent is 10:1-14: 1; the bonding method comprises the following steps:
1. placing the surfaces to be bonded of the PMMA and PDMS flexible films subjected to surface silanization treatment into oxygen or air plasma with the power of 80-300W for surface treatment for 20-90 s;
2. and spin-coating an uncured PDMS thin layer on the surface of the PDMS thin film, then attaching the PDMS thin layer to the surface of the PMMA, and performing hot-pressing bonding and curing.
The blood cell filtering and washing device provided by the invention has the following filtering and washing processes:
as shown in fig. 5, the chip 1 is placed with the sealing layer 13, the first chip 11 and the second chip 12 from top to bottom and fixed by the filter clamping member 3, the first filter clamping layer 31 directly contacts with the sealing layer 13, and the second filter clamping layer 32 directly contacts with the second chip 12.
Pour the sample into from first chip sample filtration entry according to the arrow shows, the less runner of flow resistance is independently let in to liquid, and the sample passes through the microfiltration membrane and flows out from second chip filtrate export 121, collects the outflow part waste liquid and does other detections, and second chip filtrate export 121 can be for integrated drainage tube, accelerates filtration efficiency under gravity and capillary action. After the sample is filtered, the above filtration step is repeated with buffer. The process realizes the separation of cells or impurities with different scales, the cells and the impurities with the size smaller than the target size are filtered or recovered by passing through the filter membrane, and the cells with the size larger than the target size are trapped on the filter membrane.
The filter clamping means shown in fig. 3 are removed, the chip 1 is turned 180 ° and fixed with the rinsing clamping means 4, the first rinsing clamping layer 41 being in direct contact with the closing layer 13 and the second rinsing clamping layer 42 being in direct contact with the second chip 12.
The lateral flushing holding layer 41 and the flushing liquid inlet 411 and the flushing liquid outlet 412 are closed, PBS phosphate buffer solution is injected from the flushing liquid vertical inlet 421 of the second flushing holding layer 42, and the buffer solution injected based on the flow resistance relation is filled in the lower chamber through the filter membrane 2 and then flows over the filter membrane 2. Thus, even if the filter membrane deforms downwards, the filter membrane can still be soaked in the solution. The cells on the filter membrane can fall off spontaneously under the action of gravity, and the process can last for 30s-5 min. Then, the exhaust port 422 and the flushing liquid vertical inlet 421 on the second flushing clamping layer 42 are closed, the lateral flushing liquid inlet 411 and the flushing liquid outlet 412 on the first flushing clamping layer 41 are opened, and the eluent PBS solution is introduced into the lateral flushing inlet of the chip at the flow rate of 200 mu L/min-1 mL/min, so that the method can not only realize no deposition on the filter membrane, but also ensure the integrity of the form of the spontaneously falling cells.
Fig. 7 and 8 are top views of two configurations of the microfluidic channel of the first washing nip, respectively. Wherein 411 is the lateral washing liquid inlet of the first washing clamping layer, the shape of the washing cavity 115 of the first washing clamping layer can be circular or irregular polygon, corresponding to the shape of the filter membrane opening of the chip 1, and the main purpose is to match with the washing liquid outlet, so as to reduce the dead area of the filter membrane of the chip as much as possible. To ensure the lateral flushing efficiency, the number of the flushing liquid lateral inlet channels 413 of the first flushing clamping layer is more than 2, and the arrangement mode can adopt, but is not limited to, the distribution mode shown in fig. 7 and fig. 8.
In a preferred embodiment of the present invention, the chip structure has the following dimensions:
the first chip 11 and the second chip 12 each have a thickness of 0.6mm to 3 mm. The effective area of the first chip sample filtration inlet 111 is not more than 1cm2The diameter of a flushing liquid inlet of the first flushing clamping layer is 1.5 mm-2.5 mm; width W1 of flushing liquid inlet passage of first flushing nip layer: 0.8 mm-2 mm, the depth of the channel is 100 μm-600 μm, and the diameter of the exhaust port 422 of the second flushing clamping layer is 0.2 mm-1 mm; width W2 of rinse liquid outlet passage of first rinse holding layer: 1 mm-3 mm; .
The method is described in detail as follows: after the first chip layer, the second chip layer and the filter membrane can not be bonded, the chip 1 is placed into the filtering and clamping component, and the filtering and clamping device and the chip are clamped and sealed through pressure. Pouring the sample to be processed from the upper opening to realize the filtration of the liquid in the gravity direction, and repeating the filtration step by using a buffer solution after the sample is filtered. The process realizes the separation of cells or impurities with different scales, the cells and the impurities with the size smaller than the target size penetrate through the filter membrane to be filtered, and the cells larger than the target size are intercepted on the filter membrane. And removing the pressure applied on the filtering and clamping device and the chip, overturning the chip by 180 degrees, putting the chip into a washing and clamping device, and realizing clamping and sealing of the washing and clamping device and the chip through pressure. The first washing and holding layer side washing liquid inlet 411 and the first washing and holding layer washing liquid outlet 412 are closed, PBS phosphate buffer solution is injected from the second washing and holding layer washing liquid vertical inlet 421, and the buffer solution injected based on the flow resistance relation is filled in the lower chamber through the filter membrane 2 and then overflows the filter membrane 2. Cells on the filter will fall off spontaneously under the action of gravity. The second wash and hold layer vent 422 and the second wash and hold layer vertical inlet 422 on the wash and hold member 4 are then closed, and the first wash and hold layer lateral wash inlet 411 and the first wash and hold layer wash outlet 412 are opened to allow lateral flow to transfer the cells in the chamber out of the chip.
Finally, it should be noted that: although the present invention has been described in detail with reference to the above embodiments, it should be understood by those skilled in the art that: modifications and equivalents may be made to the embodiments of the invention without departing from the spirit and scope of the invention, which is to be covered by the claims.
Claims (9)
1. A hemocyte filtration and flushing device comprises a chip, a filter membrane, a filtration clamping part and a flushing clamping part; the chip sequentially comprises a sealing layer, a first chip and a second chip; the filter membrane is positioned between the first chip and the second chip; the first chip and the second chip comprise hollow structures exposing the filter membranes; the filter clamping component comprises a first filter clamping layer in clamping contact with the closed layer and a second filter clamping layer in clamping contact with the second chip; the first filtering and clamping layer comprises a sample inlet corresponding to the filter membrane, and the second filtering and clamping layer comprises a filtrate outlet corresponding to the filter membrane; the flushing clamping component comprises a first flushing clamping layer in clamping contact with the sealing layer and a second flushing clamping layer in clamping contact with the second chip; the first flushing clamping layer comprises a lateral flushing liquid inlet corresponding to one side of the filter membrane; the first washing clamping layer comprises a washing liquid outlet corresponding to the position on the other side of the filter membrane, the second washing clamping layer comprises a vertical washing liquid inlet corresponding to the position above the filter membrane and an exhaust port, and the first washing clamping layer is characterized in that: the first flushing clamping layer comprises a flushing liquid flow channel which is communicated with the lateral flushing liquid inlet and the flushing liquid outlet; the filter membrane is irreversibly bonded with the first chip and the second chip.
2. The hemocyte filtration irrigation device of claim 1 wherein: the thickness of the first chip and the second chip is 0.6mm-3 mm; the effective area of the filter membrane corresponding to the sample inlet and the sample outlet is less than 1cm2(ii) a The diameters of the lateral flushing liquid inlet, the vertical flushing liquid inlet and the flushing liquid outlet are 1.5 mm-2.5 mm; the diameter of the exhaust port is 0.2 mm-1 mm.
3. The hemocyte filtration and washing device of claim 1 wherein: the flushing liquid flow channel and the filter membrane are positioned on different planes.
4. The hemocyte filtration irrigation device of claim 3 wherein: first filtration centre gripping layer is including closed valve, it is regional to be provided with the valve on the flush fluid runner, the regional diameter of valve does 1.1~2 times of flush fluid runner width, the direction of depth be with closed valve curved surface complex arc curved surface.
5. The hemocyte filtration and washing device of claim 4 wherein: the flushing fluid flow passage comprises a flushing fluid inlet flow passage connected with the lateral flushing fluid inlet and a flushing fluid outlet flow passage connected with the flushing fluid outlet.
6. The hemocyte filtration and washing device of claim 5 wherein: the washing liquid inlet flow passage is 2 or more, the washing liquid outlet flow passage is one, and the width ratio of the washing liquid outlet flow passage to the washing liquid inlet flow passage is the inverse ratio of the number of the washing liquid outlet flow passage to the number of the washing liquid inlet flow passages.
7. The hemocyte filtration and washing device of claim 6 wherein: the irreversible bond comprises a thermocompression bond.
8. The hemocyte filtration irrigation device of claim 7 wherein: the first chip and the second chip are PMMA, the thermocompression bonding treatment comprises the steps of placing the filter membrane material between the first chip and the second chip, fixing the filter membrane material, placing the filter membrane material into an ethanol or acetone solvent, and carrying out ultrasonic treatment on the filter membrane material by 100-360W for no more than 3 minutes at the temperature lower than 45 ℃.
9. The hemocyte filtration and washing device of claim 7 wherein: the first chip and the second chip are PMMA, and are respectively bonded by using a thermal compression bonding method with the thickness of 256nm and the thickness of 0.1J/cm before thermal compression bonding2-10 J/cm2The first and second chips are irradiated with ultraviolet light.
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| CN201911083013.9A CN112774742B (en) | 2019-11-07 | 2019-11-07 | Blood cell filters washing unit |
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| CN201911083013.9A CN112774742B (en) | 2019-11-07 | 2019-11-07 | Blood cell filters washing unit |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN205856486U (en) * | 2015-12-07 | 2017-01-04 | 内江师范学院 | A kind of easy micro-fluidic chip |
| CN108499619A (en) * | 2018-03-09 | 2018-09-07 | 复旦大学 | A kind of integrated micro-fluidic filtrating chip of film and its preparation method and application |
| CN109633174A (en) * | 2019-01-09 | 2019-04-16 | 昆明医科大学海源学院 | A kind of super quick detection method of c reactive protein based on biotin quantum dot probe |
| WO2019180662A1 (en) * | 2018-03-22 | 2019-09-26 | Clinical Sensors, Inc. | Sensing devices and methods of production |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100261159A1 (en) * | 2000-10-10 | 2010-10-14 | Robert Hess | Apparatus for assay, synthesis and storage, and methods of manufacture, use, and manipulation thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN205856486U (en) * | 2015-12-07 | 2017-01-04 | 内江师范学院 | A kind of easy micro-fluidic chip |
| CN108499619A (en) * | 2018-03-09 | 2018-09-07 | 复旦大学 | A kind of integrated micro-fluidic filtrating chip of film and its preparation method and application |
| WO2019180662A1 (en) * | 2018-03-22 | 2019-09-26 | Clinical Sensors, Inc. | Sensing devices and methods of production |
| CN109633174A (en) * | 2019-01-09 | 2019-04-16 | 昆明医科大学海源学院 | A kind of super quick detection method of c reactive protein based on biotin quantum dot probe |
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