CN105462812A - Preparation method of cell capturing and releasing chip based on paraffin substrate film - Google Patents
Preparation method of cell capturing and releasing chip based on paraffin substrate film Download PDFInfo
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- CN105462812A CN105462812A CN201510922070.7A CN201510922070A CN105462812A CN 105462812 A CN105462812 A CN 105462812A CN 201510922070 A CN201510922070 A CN 201510922070A CN 105462812 A CN105462812 A CN 105462812A
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- 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
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- 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
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Abstract
The invention relates to a preparation method of a chip realizing rapid cell capturing and releasing based on a paraffin substrate film. The preparation method comprises the steps of substrate selection, etching patterns embedding, wet etching and substrate bonding during the chip preparation. On the basis of wet etching, paraffin is melted on a substrate after wet etching, meanwhile the surface facing upwards of a clean substrate is uniformly coated with edible sesame oil, the substrate with edible sesame oil is placed on a heating table to be heated, then the surface with the edible sesame oil of the substrate coated with the edible sesame oil covers the substrate with paraffin in an aligning manner, thus the preparation of the paraffin film is completed, and the cell capturing and releasing chip based on the paraffin substrate film is obtained. The problem that the existing micro-fluidic chip only can capture cells but can not release the cells is solved, the amount of the needed material is small, a processing method is simple, and the prepared chip has very high application value and economic value in the field of medical analysis.
Description
Technical field
The present invention relates to the preparation method of micro-fluidic chip, specifically a kind of cell capture based on paraffin substrate film and the preparation method discharging chip.
Background technology
In recent years, micro-fluidic chip has become the focus of cellular segregation research, the fast development of micro-fluidic chip and applying, and provides great convenience to analysis field.CTCs(CirculatingTumorCells circulation of blood tumour cell in blood of human body) content very low, this is concerning existing method for separating and analyzing being a huge challenge.Also do not have other good method script quantity in blood well can be separated with regard to few CTCs at present, high-sensitive detection counting can not be carried out even if can separate and carry out follow-up a series of biochemical analysis.The appearance that at this time CTCs(catches chip can greatly address this problem, and the appearance of cell capture chip has great meaning for the quick discovery of CTCs and early stage Clinics and Practices.
But the newest research results about CTCs chip of reporting for work at present, although it is higher to the capture rate of CTCs, the CTCs captured is difficult to discharge, and what inconvenience was follow-up accompanies supporting grade for biochemical analysis again.If public announcement of a patent application number is for CN103235124A and public announcement of a patent application number are for as described in CN104324768A, although they can carry out catching of cell and capture rate is higher, but its cell captured can not effectively discharge, inconvenient follow-up enlarged culturing and analyzing again.
Summary of the invention
The object of the invention is to overcome above-mentioned the deficiencies in the prior art, provide the preparation method of a kind of cell capture based on paraffin substrate film with release chip, low cost, processing is simple.
Another object of the present invention is to provide a kind of paraffin substrate film chip that adopts to catch the method with release cells, catches with release efficiency high.
To achieve these goals, the present invention solves the problems of the technologies described above adopted technical scheme and is:
Based on the cell capture of paraffin substrate film and a preparation method for release chip, concrete steps are as follows:
(1) selection of substrate and cleaning: choose glass, quartz or silicon as substrate, the two panels substrate chosen cleaned, removes the impurity such as the greasy dirt of substrate surface;
(2) etch mask process: in the baking oven of 50 DEG C ~ 100 DEG C, drying and processing is carried out to cleaned substrate, adhere to the mask of chemical resistance in the upper and lower surface of two panels substrate after drying and processing, pass through laser cutting, the area part that namely will etch is cut out, remove the protection against corrosion mask cut out, obtain the region that will etch;
The object of mask process is to expose the part needing to carry out the substrate etched, and other parts of the substrate not needing to carry out to etch are protected.
(3) wet etching: by the substrate handled well upper slice, vertically to be put in etching liquid slowly according to the speed of 0.01ml/min-1mm/min by fixture and etch, the upper slice groove floor that etches is slope, the substrate bottom sheet handled well is directly immersed in etching liquid and etches, the groove floor level that bottom sheet etches;
(4) punch: to get rid of slice and the etch mask on bottom sheet surface, make complete out exposed of whole substrate, after clean is carried out to substrate, punch to upper slice, in the punching of the region of at least two different contour planes of upper, two holes are respectively ingate and outlet opening.
(5) preparation of Parafilm: cut the groove that a small amount of paraffin is placed in bottom sheet, bottom sheet is placed in thermal station and is heated to 55 DEG C, now paraffin melts completely, grease is spread upon uniformly a slice clean without etching substrate on and heat, the substrate scribbling grease is covered slowly and has in the bottom sheet of paraffin in thawing, shift out from thermal station slowly, allow the chip slow cooling combined until the paraffin in bottom sheet solidifies completely, then the chip combined is taken apart, get rid of the paraffin that bottom sheet groove periphery is unnecessary, complete the preparation of bottom sheet Parafilm.
The thickness of described Parafilm is 0.1-1.5mm, and namely after bottom sheet etching, the height of its etch areas is 0.1-1.5mm.
(6) upper slice with the combination of bottom sheet: choose PDMS(polydimethylsiloxane) thin slice to bottom sheet and upper slice carry out bonding, just obtain the cell capture based on paraffin substrate film and release chip.
Adopt the chip based on paraffin substrate film to carry out cell capture and a method for releasing, its step is as follows:
(1) chip with paraffin substrate film of preparation is placed in fixture, with the speed of 0.01ul/min-1ul/min, cultured cell is injected into bonding completely chip from ingate by syringe pump, after continuing to pass into the regular hour, cell is stuck in etched volume, and biological tissue's buffered soln flows out from outlet opening.
(2) the Parafilm laser of chip is irradiated or is placed in thermal station heat, at this time part paraffin starts to melt, meanwhile syringe pump is started working and inject PBS damping fluid (phosphate buffered saline(PBS)) in chip, cell above the Parafilm melted just to be rushed out, achieves the release to cell.
Beneficial effect: the present invention changes the problem that the micro flow chip in the past produced can not discharge the cell captured.Preparation method of the present invention, complete processing is simple, and the chip prepared can discharge the cell captured, and has wide market outlook and marketable value, can be widely used in analysis field.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of chip.Wherein 1(a) be upper slice schematic diagram, trapezoidal peripheral regions is overlay film region, 1(b) and be bottom sheet schematic diagram, little trapezoidal surrounding is overlay film region.
Fig. 2 is the cell capture efficiency under different injection rate.
Embodiment
Below in conjunction with embodiment, the present invention is made further instructions in cell capture and release.Etching liquid is typical buffer oxide silicon etching liquid, wherein fluorinated ammonium, and (BOE:BufferOxideEtcher) and deionized water are blent by the mass ratio of 1:10 and formed.
embodiment 1
Adopt the chip based on paraffin substrate film to carry out catching of cell and a method for releasing, concrete steps are as follows:
1. choice of the substrates is glass, wherein the length of substrate of glass is 75mm, width is 25mm, thickness is 2mm, the two panels substrate chosen is cleaned under the deionized water of flowing, removes the impurity such as the greasy dirt of substrate surface, in 50-100 DEG C of baking oven, drying and processing is carried out to cleaned substrate.
2, the protection gummed paper of chemical resistance is pasted in the upper and lower surface of substrate; the protection gummed paper of upper cuts out 1(a in as Fig. 1 with laser engraving machine) as described in trapezoid area; trapezoidal upper base is 9mm; go to the bottom as 18mm; the height of whole trapezoid area is 60mm; bottom sheet cuts out as 1(b in Fig. 1 with laser engraving machine) as described in the trapezoidal upper base of little trapezoid area be 14mm; go to the bottom as 18mm; the height of whole trapezoid area is 20mm; tear upper slice and the protection gummed paper of bottom sheet cutting zone, obtain the region that will etch.
3, by the substrate handled well upper slice, vertically to be put into slowly in etching liquid according to the speed of 1mm/min by fixture and carry out etching 75min, bottom sheet is directly immersed in etching liquid and etches, and after 30min bottom sheet etching, the height of its etch areas is 1.5mm.
4, to get rid of slice and the etch mask on bottom sheet surface, make complete out exposed of whole substrate, after clean is carried out to substrate, punch to upper slice, punch in the region of at least two different contour planes of upper, two holes are respectively ingate and outlet opening, and processing the diameter portalled is 0.5-1mm.
5, cut the groove that a small amount of paraffin is placed in bottom sheet, be placed in 55 DEG C of thermal station and heat, until paraffin is melted in the groove of bottom sheet completely; Meanwhile edible sesame oil is spread upon uniformly clean without a slice substrate of etching is placed in thermal station and heats.
6, the substrate scribbling edible sesame oil is covered slowly on the bottom sheet that thawing has paraffin, shift out from thermal station slowly, allow the substrate slow cooling combined until melt and have the paraffin in the bottom sheet of paraffin to solidify completely, then rapid the substrate combined to be taken apart, get rid of with the unnecessary paraffin of the groove periphery of bottom sheet, the substrate scribbling edible sesame oil is cleaned up simultaneously, complete the preparation of the substrate with Parafilm.
7, choose PDMS(polydimethylsiloxane) to through above-mentioned process upper slice and bottom sheet carry out bonding, just obtain the catching and discharging chip of the rapid cellular based on paraffin substrate film that the present invention needs to prepare.
8, the chip prepared is placed in fixture, cultured breast cancer cell and prostate gland cancerous tumor cell are injected 1 hour to bonding completely chip with the speed of 1ul/min from ingate by syringe pump, breast cancer cell is stuck in etched volume, biological tissue's buffered soln flows out from outlet opening, completes catching cell.
9, the Parafilm part laser of chip is irradiated, at this time part paraffin starts to melt, meanwhile syringe pump is started working and inject PBS damping fluid in chip, and cell above the Parafilm being irradiated with a laser part is just rushed out, achieves the release to cell.
embodiment 2
Adopt the chip based on paraffin substrate film to carry out catching of cell and a method for releasing, concrete steps are as follows:
1. choice of the substrates is glass, wherein the length of substrate of glass is 75mm, width is 25mm, thickness is 2mm, the two panels substrate chosen is cleaned under the deionized water of flowing, removes the impurity such as the greasy dirt of substrate surface, in 50-100 DEG C of baking oven, drying and processing is carried out to cleaned substrate.
2, the protection gummed paper of chemical resistance is pasted in the upper and lower surface of substrate; the protection gummed paper of upper cuts out 1(a in as Fig. 1 with laser engraving machine) as described in the trapezoidal upper base of trapezoid area be 9mm; go to the bottom as 18mm; the height of whole trapezoid area is 45mm; bottom sheet cuts out as 1(b in Fig. 1 with laser engraving machine) as described in the trapezoidal upper base of little trapezoid area be 9mm; go to the bottom as 18mm, the height of whole trapezoid area is 18mm
Tear upper slice and the protection gummed paper of bottom sheet cutting zone, obtain the region that will etch.
3, by the substrate handled well upper slice, vertically to be put into slowly in etching liquid according to the speed of 0.5mm/min by fixture and carry out etching 70min, the substrate that bottom sheet is handled well is directly immersed in etching liquid and etches, and after etching 25min bottom sheet etching, the height of its etch areas is 1.25mm.
4, to get rid of slice and the etch mask on bottom sheet surface, make complete out exposed of whole substrate, after clean is carried out to substrate, punch to upper slice, punch in the region of at least two different contour planes of upper, two holes are respectively ingate and outlet opening, and processing the diameter portalled is 0.5-1mm.
5, cut the groove that a small amount of paraffin is placed in bottom sheet, be placed in 55 DEG C of thermal station and heat, until paraffin is melted in the groove of bottom sheet completely; Meanwhile edible sesame oil is spread upon uniformly clean without a slice substrate of etching is placed in thermal station and heats.
6, the substrate scribbling edible sesame oil is covered slowly on the bottom sheet that thawing has paraffin, shift out from thermal station slowly, allow the substrate slow cooling combined until melt and have the paraffin in the bottom sheet of paraffin to solidify completely, then rapid the substrate combined to be taken apart, get rid of with the unnecessary paraffin of the groove periphery of bottom sheet, the substrate scribbling edible sesame oil is cleaned up simultaneously, complete the preparation of the substrate with Parafilm.
7, choose PDMS(polydimethylsiloxane) to through above-mentioned process upper slice and bottom sheet carry out bonding, just obtain the catching and discharging chip of the rapid cellular based on paraffin substrate film that the present invention needs to prepare.
8, the chip prepared is placed in fixture, cultured breast cancer cell and prostate gland cancerous tumor cell are injected 1 hour to bonding completely chip with the speed of 0.75ul/min from ingate by syringe pump, prostate gland cancerous tumor cell is stuck in etched volume, biological tissue's buffered soln flows out from outlet opening, completes catching cell
9, be placed in thermal station by whole for chip, meanwhile syringe pump is started working and inject PBS damping fluid in chip, is all discharged by the cell captured, and so just achieves the object of the cell captured being carried out to all release.
embodiment 3
Adopt the chip based on paraffin substrate film to carry out catching of cell and a method for releasing, concrete steps are as follows:
1. choice of the substrates is quartz, wherein the length of quartz substrate is 75mm, width is 25mm, thickness is 2mm, the two panels substrate chosen is cleaned under the deionized water of flowing, removes the impurity such as the greasy dirt of substrate surface, in 50-100 DEG C of baking oven, drying and processing is carried out to cleaned substrate.
2, the protection gummed paper of chemical resistance is pasted in the upper and lower surface of substrate; the protection gummed paper of upper and bottom sheet cuts out with laser engraving machine length is 40mm, width is the rectangle of 10mm; tear upper slice and the protection gummed paper of bottom sheet cutting zone, obtain the region that will etch.
3, by the substrate handled well upper slice, vertically to be put into slowly in etching liquid according to the speed of 0.8mm/min by fixture and carry out etching 65min, the substrate that bottom sheet is handled well is directly immersed in etching liquid and carries out etching 20min, and after bottom sheet etching, the height of its etch areas is 1mm.
4, to get rid of slice and the etch mask on bottom sheet surface, make complete out exposed of whole substrate, after clean is carried out to substrate, punch to upper slice, punch in the region of at least two different contour planes of upper, two holes are respectively ingate and outlet opening, and processing the diameter portalled is 0.5-1mm.
5, cut the groove that a small amount of paraffin is placed in bottom sheet, be placed in 55 DEG C of thermal station and heat, until paraffin is melted in the groove of bottom sheet completely; Meanwhile edible sesame oil is spread upon uniformly clean without a slice substrate of etching is placed in thermal station and heats.
6, the substrate scribbling edible sesame oil is covered slowly on the bottom sheet that thawing has paraffin, shift out from thermal station slowly, allow the substrate slow cooling combined until melt and have the paraffin in the bottom sheet of paraffin to solidify completely, then rapid the substrate combined to be taken apart, get rid of with the unnecessary paraffin of the groove periphery of bottom sheet, the substrate scribbling edible sesame oil is cleaned up simultaneously, complete the preparation of the substrate with Parafilm.
7, choose PDMS(polydimethylsiloxane) to through above-mentioned process upper slice and bottom sheet carry out bonding, just obtain the catching and discharging chip of the rapid cellular based on paraffin substrate film that the present invention needs to prepare.
8, the chip prepared is placed in fixture, cultured breast cancer cell and prostate gland cancerous tumor cell are injected 1 hour to bonding completely chip with the speed of 0.5ul/min from ingate by syringe pump, mammary cancer breast cancer cell and prostate gland cancerous tumor cell are stuck in etched volume, biological tissue's buffered soln flows out from outlet opening, completes catching cell
9, be placed in thermal station by whole for chip, meanwhile syringe pump is started working and inject PBS damping fluid in chip, is all discharged by the cell captured, and so just achieves the object of the cell captured being carried out to all release.
embodiment 4
Adopt the chip based on paraffin substrate film to carry out catching of cell and a method for releasing, concrete steps are as follows:
1. choice of the substrates is quartz, wherein the length of quartz substrate is 75mm, width is 25mm, thickness is 2mm, the two panels substrate chosen is cleaned under the deionized water of flowing, removes the impurity such as the greasy dirt of substrate surface, in 50-100 DEG C of baking oven, drying and processing is carried out to cleaned substrate.
2, the protection gummed paper of chemical resistance is pasted in the upper and lower surface of substrate; the protection gummed paper of upper and bottom sheet cuts out with laser engraving machine length is 30mm, width is the rectangular etch areas of 15mm; tear upper slice and the protection gummed paper of bottom sheet cutting zone, obtain the region that will etch.
3, by the substrate handled well upper slice, vertically to be put into slowly in etching liquid according to the speed of 0.1mm/min by fixture and carry out etching 60min, the substrate that bottom sheet is handled well is directly immersed in etching liquid and carries out etching 18min, and after bottom sheet etching, the height of its etch areas is 0.9mm.
4, to get rid of slice and the etch mask on bottom sheet surface, make complete out exposed of whole substrate, after clean is carried out to substrate, punch to upper slice, punch in the region of at least two different contour planes of upper, two holes are respectively ingate and outlet opening, and processing the diameter portalled is 0.5-1mm.
5, cut the groove that a small amount of paraffin is placed in bottom sheet, be placed in 55 DEG C of thermal station and heat, until paraffin is melted in the groove of bottom sheet completely; Meanwhile edible sesame oil is spread upon uniformly clean without a slice substrate of etching is placed in thermal station and heats.
6, the substrate scribbling edible sesame oil is covered slowly on the bottom sheet that thawing has paraffin, shift out from thermal station slowly, allow the substrate slow cooling combined until melt and have the paraffin in the bottom sheet of paraffin to solidify completely, then rapid the substrate combined to be taken apart, get rid of with the unnecessary paraffin of the groove periphery of bottom sheet, the substrate scribbling edible sesame oil is cleaned up simultaneously, complete the preparation of the substrate with Parafilm.
7, choose PDMS(polydimethylsiloxane) to through above-mentioned process upper slice and bottom sheet carry out bonding, just obtain the catching and discharging chip of the rapid cellular based on paraffin substrate film that the present invention needs to prepare.
8, the chip prepared is placed in fixture, cultured breast cancer cell and prostate gland cancerous tumor cell are injected 1 hour to bonding completely chip with the speed of 0.25ul/min from ingate by syringe pump, breast cancer cell and prostate gland cancerous tumor cell are stuck in etched volume, biological tissue's buffered soln flows out from outlet opening, completes catching cell
9, be placed in thermal station by whole for chip, meanwhile syringe pump is started working and inject PBS damping fluid in chip, is all discharged by the cell captured, and so just achieves the object of the cell captured being carried out to all release.
embodiment 5
Adopt the chip based on paraffin substrate film to carry out catching of cell and a method for releasing, concrete steps are as follows:
1. choice of the substrates is silicon chip, length wherein at the bottom of silicon wafer-based is 75mm, width is 25mm, thickness is 2mm, the two panels substrate chosen is cleaned under the deionized water of flowing, removes the impurity such as the greasy dirt of substrate surface, in 50-100 DEG C of baking oven, drying and processing is carried out to cleaned substrate.
2, the protection gummed paper of chemical resistance is pasted in the upper and lower surface of substrate; the protection gummed paper of upper and bottom sheet cuts out with laser engraving machine length is 30mm, width is the rectangular etch areas of 15mm; tear upper slice and the protection gummed paper of bottom sheet cutting zone, obtain the region that will etch.
3, by the substrate handled well upper slice, vertically to be put into slowly in etching liquid according to the speed of 0.1mm/min by fixture and carry out etching 60min, the substrate that bottom sheet is handled well is directly immersed in etching liquid and carries out etching 10min, and after bottom sheet etching, the height of its etch areas is 0.5mm.
4, to get rid of slice and the etch mask on bottom sheet surface, make complete out exposed of whole substrate, after clean is carried out to substrate, punch to upper slice, punch in the region of at least two different contour planes of upper, two holes are respectively ingate and outlet opening, and processing the diameter portalled is 0.5-1mm.
5, cut the groove that a small amount of paraffin is placed in bottom sheet, be placed in 55 DEG C of thermal station and heat, until paraffin is melted in the groove of bottom sheet completely; Meanwhile edible sesame oil is spread upon uniformly clean without a slice substrate of etching is placed in thermal station and heats.
6, the substrate scribbling edible sesame oil is covered slowly on the bottom sheet that thawing has paraffin, shift out from thermal station slowly, allow the substrate slow cooling combined until melt and have the paraffin in the bottom sheet of paraffin to solidify completely, then rapid the substrate combined to be taken apart, get rid of with the unnecessary paraffin of the groove periphery of bottom sheet, the substrate scribbling edible sesame oil is cleaned up simultaneously, complete the preparation of the substrate with Parafilm.
7, choose PDMS(polydimethylsiloxane) to through above-mentioned process upper slice and bottom sheet carry out bonding, just obtain the catching and discharging chip of the rapid cellular based on paraffin substrate film that the present invention needs to prepare.
8, the chip prepared is placed in fixture, cultured cell breast cancer cell and prostate gland cancerous tumor cell are injected 1 hour to bonding completely chip with the speed of 0.01ul/min from ingate by syringe pump, breast cancer cell and prostate gland cancerous tumor cell are stuck in etched volume, biological tissue's buffered soln flows out from outlet opening, completes catching cell.
9, be placed in thermal station by whole for chip, meanwhile syringe pump is started working and inject PBS damping fluid in chip, is all discharged by the cell captured, and so just achieves the object of the cell captured being carried out to all release.
As shown in Figure 2, in certain injection length, injection speed its capture rate lower is higher, and capture rate is up to 95%.Observed by microscope by the cell that aforesaid method is captured, find that the cellularstructure captured is complete, without disrepair phenomenon.The ratio injecting the cell quantity of formed objects when the cell capture efficiency that the present invention adopts is and is started with experiment by the cell quantity finally stopped in the chips calculates the capture rate of cell.
The present invention changes the problem that the micro flow chip in the past produced can not discharge the cell captured.Preparation method of the present invention, complete processing is simple, and the chip prepared can discharge the cell captured, and has wide market outlook and marketable value, can be widely used in analysis field.
Claims (3)
1., based on the cell capture of paraffin substrate film and a preparation method for release chip, its step is as follows:
(1) selection of substrate and cleaning: choose glass, quartz or silicon as substrate, the two panels substrate chosen cleaned, removes the impurity such as the greasy dirt of substrate surface;
(2) etch mask process: in the baking oven of 50 DEG C ~ 100 DEG C, drying and processing is carried out to cleaned substrate, adhere to the mask of chemical resistance in the upper and lower surface of two panels substrate after drying and processing, pass through laser cutting, the area part that namely will etch is cut out, remove the protection against corrosion mask cut out, obtain the region that will etch;
(3) wet etching: by the substrate handled well upper slice, vertically to be put in etching liquid slowly according to the speed of 0.01ml/min-1mm/min by fixture and etch, the upper slice groove floor that etches is slope, the substrate bottom sheet handled well is directly immersed in etching liquid and etches, the groove floor level that bottom sheet etches;
(4) punch: to get rid of slice and the etch mask on bottom sheet surface, make complete out exposed of whole substrate, after clean is carried out to substrate, punch to upper slice, in the punching of the region of at least two different contour planes of upper, two holes are respectively ingate and outlet opening;
(5) preparation of Parafilm: cut the groove that a small amount of paraffin is placed in bottom sheet, bottom sheet is placed in thermal station and is heated to 55 DEG C, now paraffin melts completely, grease is spread upon uniformly a slice clean without etching substrate on and heat, the substrate scribbling grease is covered slowly and has in the bottom sheet of paraffin in thawing, shift out from thermal station slowly, allow the chip slow cooling combined until the paraffin in bottom sheet solidifies completely, then the chip combined is taken apart, get rid of the paraffin that bottom sheet groove periphery is unnecessary, complete the preparation of bottom sheet Parafilm,
(6) upper slice with the combination of bottom sheet: choose PDMS thin slice to bottom sheet and upper slice carry out bonding, just obtain the cell capture based on paraffin substrate film and release chip.
2. the cell capture based on paraffin substrate film according to claim 1 and the preparation method of release chip, is characterized in that: the thickness of described Parafilm is 0.1-1.5mm.
3. adopt chip according to claim 1 to catch the method with release cells, its step is as follows:
(1) chip with paraffin substrate film of preparation is placed in fixture, with the speed of 0.01ul/min-1ul/min, cultured cell is injected into bonding completely chip from ingate by syringe pump, after continuing to pass into the regular hour, cell is stuck in etched volume, and biological tissue's buffered soln flows out from outlet opening;
(2) the Parafilm laser in chip is irradiated or is directly placed in thermal station heat, at this time part paraffin starts to melt, meanwhile syringe pump is started working and inject PBS damping fluid in chip, cell above the Parafilm melted just to be rushed out, achieves the release to cell.
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| CN110108623A (en) * | 2019-04-30 | 2019-08-09 | 武汉纺织大学 | A kind of greasy dirt grain testing apparatus and method based on micro-fluidic chip |
| CN114042478A (en) * | 2021-11-11 | 2022-02-15 | 常州大学 | Photoresponse transverse filtering chip and separation and release method of single cancer cell |
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