CN113637580A - Multi-force field coupled cell tissue culture chip - Google Patents
Multi-force field coupled cell tissue culture chip Download PDFInfo
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- CN113637580A CN113637580A CN202010346127.4A CN202010346127A CN113637580A CN 113637580 A CN113637580 A CN 113637580A CN 202010346127 A CN202010346127 A CN 202010346127A CN 113637580 A CN113637580 A CN 113637580A
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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
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/40—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pressure
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
Abstract
The invention belongs to the technical field of biology, and relates to a multi-force field coupled cell tissue culture chip, in particular to a cell tissue culture chip for multi-force field coupling of mechanical mechanics, hydromechanics and hydrostatic pressure. In the chip, the air cylinder is connected with the air pressure control system, the chip is connected with the hydraulic control system, and the chip is driven to stretch and pressurize inside the chip by the control system operating the switch of the electromagnetic valve and the voltage applying time. The invention has simple and convenient operation, can realize dynamic culture of cell tissues and can apply multi-force field stimulation simultaneously, and is suitable for the field of cell tissue mechanics.
Description
Technical Field
The invention belongs to the technical field of biology, relates to a multi-force-field coupled cell tissue culture chip, and particularly relates to a multi-force-field coupled cell tissue culture chip for mechanical mechanics, hydromechanics and hydrostatic pressure.
Background
The traditional cell tissue culture technology comprises the steps of performing static two-dimensional culture on cell tissues by using a culture dish, a culture bottle, a pore plate and the like; however, the microenvironment of the cell tissue is complex, which is a complex set with multi-factor composition and space-time variability, and plays a decisive role in the behavior and the function of the cell tissue, and research practices show that the traditional static culture method is difficult to provide a real microenvironment for the cell tissue in vitro, so that the research result of the in vitro cell tissue biology is far from the in vivo situation. With the development of the micro-fluidic chip technology, the combination of the micro-fluidic chip technology and the cell tissue culture technology provides possibility for the simulation of the cell tissue microenvironment, can accurately control the key parameters of the cell tissue survival microenvironment, and can observe the response of the cell tissue to the parameter change in the microenvironment in real time.
The microfluidic chip is an application technology for controlling fluid in micrometer scale and space, and has a function of reducing the basic functions of laboratories of chemistry, biology, physics and the like to a chip with a volume of only a few square centimeters, and is also called a Lab-on-chip (Lab-on-chip). Compared with other platforms, the microfluidic chip has the characteristics of low material cost, low reagent consumption, high reaction speed, high sensitivity, good portability, simple operation, flexible composition and scale integration of various unit technologies on a micro-controllable platform and the like. In addition, the microfluidic chip is usually made of PDMS, which has characteristics of light transmittance, air permeability, biocompatibility, ductility and the like, and is widely applied to the fields of biochemistry, molecular biology and the like.
At present, research reports of multi-force field coupled cell tissue culture chips aiming at mechanical mechanics, hydromechanics and hydrostatic pressure are rarely found at home and abroad.
Based on the research foundation and the current situation of the prior art, the inventor of the present application intends to provide a novel multi-force field coupled cell tissue culture chip, in particular a multi-force field coupled cell tissue culture chip for use in mechanics, hydromechanics and hydrostatic pressure.
References relevant to the present invention are:
[1]Tumor Microenvironment:Interactions and Therapy[J].Journal of Cellular Physiology,2019,234(5).
[2]Sasha H.Bakhru,Christopher Highley,Stefan Zappe.Application of Microfluidics in Stem Cell and Tissue Engineering[M].Microfluidic Technologies for Human Health.2013.
[3] lin propylic bearing microfluidic chip lab [ J ] 2003.
Disclosure of Invention
The invention aims to provide a novel multi-force-field coupled cell tissue culture chip based on the research foundation and the current situation of the prior art, in particular to a multi-force-field coupled cell tissue culture chip for mechanical mechanics, hydromechanics and hydrostatic pressure. The invention can realize the scheme of cell tissue culture of independent application of mechanical mechanics, hydromechanics and hydrostatic pressure or coupling of multiple force fields, realize the controllable reappearance of the in vitro cell tissue microenvironment, and provide a powerful technical platform for the research of the interaction of the cell tissue and the microenvironment.
The purpose of the invention is realized by the following technical scheme:
a multi-force field coupled cell tissue culture chip is characterized in that,
the chip comprises upper and lower layer PDMS chip, inlet, liquid outlet, three hole chip anchor clamps and the parallel pneumatic finger cylinder of mutual bonding, and upper and lower layer PDMS chip is the cuboid of equidimension to laminate each other through the plasma bonding mode, set up various opening decurrent recess runners, cell tissue culture microcavity and screw through-hole on the last binding face of lower layer PDMS chip. An inlet through hole, an outlet through hole and a screw through hole which correspond to the groove flow channel of the lower-layer chip are formed in the upper-layer PDMS chip; the three-hole chip clamp is a cuboid made of acrylic materials, and through holes of the three-hole chip clamp are formed by using a laser engraving machine; the fixture is used for fixing a multi-force field coupling cell tissue culture chip and a parallel pneumatic finger cylinder; after the assembly is finished, the air cylinder is connected with the air pressure control system, the chip is connected with the hydraulic control system, and the control system controls the switch of the electromagnetic valve and the voltage application time to drive the stretching movement of the chip and the internal pressurization of the chip.
The invention relates to a multi-force field coupled cell tissue culture chip, which is used for mechanical mechanics, hydromechanics and hydrostatic force multi-force field coupling.
In the invention, after the upper PDMS chip (2) and the lower PDMS chip (1) are bonded, the luer connector is screwed into the inlet through hole (11) and the outlet through hole (10), other reagents such as cell tissue suspension, cell tissue culture medium and the like are filled into the round-corner square cell tissue culture microcavity (4) from the luer connector screwed into the inlet through hole (11), redundant gas in the microcavity is discharged through the luer connector screwed into the outlet through hole (10), and the luer connector screwed into the inlet through hole (11) is connected with a hydraulic control system;
in the invention, the upper PDMS chip (2) is a cuboid with the thickness of 88.5mm multiplied by 40mm multiplied by 4mm, the inlet (11) and outlet through holes (10) are through holes with the thickness d equal to 2mm, and the screw through holes (9) are round through holes with the thickness d equal to 3 mm;
in the invention, the lower PDMS chip (1) is a 88.5mm × 40mm × 4mm cuboid, a cuboid groove with a 22mm × 20mm × 1.7mm fillet (R ═ 1mm) is downwards formed on the cell tissue culture micro-cavity, the liquid flow channel (7) is formed by an arc with a depth of 1.7mm (R ═ 6.2 mm), the liquid inlet cavity (6) and the liquid outlet cavity (5) are circular with a depth of 1.7mm (R ═ 2 mm), the screw through hole (8) is a circular through hole with a depth of 3mm, and the center of the screw through hole is aligned with the center of the clamp fixing through hole (9) of the upper PDMS chip (2);
according to the invention, the three-hole chip clamp (3) is a 40mm × 15mm × 4mm acrylic cuboid, a laser engraving machine is utilized to form a circular through hole with d ═ 3.5mm in the three-hole chip clamp (3), and the circle center of the circular through hole is aligned with the circle centers of the screw through holes (9) and (8) of the upper-layer PDMS chip (2) and the lower-layer PDMS chip (1) and the threaded hole of the horizontal pneumatic finger cylinder;
in the chip, the bonding mode of the upper layer (2) and the lower layer PDMS chip (1) is plasma irreversible bonding;
the upper-layer PDMS chip (2) and the lower-layer PDMS chip (1) are fixed on a parallel pneumatic finger cylinder sliding platform by using an M3 stainless steel screw and a three-hole chip clamp (3);
the chip can realize a cell tissue culture scheme of independent application of mechanical force, hydrodynamics and hydrostatic pressure or coupling of multiple force fields, realize controllable reproduction of in-vitro cell tissue microenvironment, and can observe the response of cell tissues to parameter changes in the microenvironment in real time by combining the technologies of microscope observation, immunoassay and the like.
Compared with the prior art, the invention has the beneficial effects that:
1. the multi-force field coupled cell tissue culture chip can realize the single application of mechanical mechanics, hydromechanics and hydrostatic pressure or the multi-force field coupled cell tissue culture scheme, and is convenient for the research of the cell tissue mechanics aiming at single-factor change or multi-factor change;
2. the multi-force field coupled cell tissue culture chip can accurately control key parameters of a cell tissue survival microenvironment;
3. the multi-force-field coupled cell tissue culture chip can observe the response of cell tissues to parameter changes in a microenvironment in real time;
4. the multi-force field coupled cell tissue culture chip has good robustness and can be used for researching cell tissue mechanics experiments for a long time.
Drawings
FIG. 1 is a schematic diagram of a multi-force field coupled cell tissue culture chip,
wherein, 1, a lower PDMS chip; 2, an upper PDMS chip; 3, a round-corner rectangular cell tissue culture chamber; 4, a liquid inlet cavity; 5, a liquid outlet cavity; 6, a liquid flow channel; 7, a lower chip screw through hole; 8, an upper layer chip screw through hole; 9, liquid inlet through holes; and 10, liquid outlet through holes.
FIG. 2 is a schematic view of a three-hole chip clamp.
Fig. 3 is a parallel pneumatic finger cylinder, wherein, 1, a sliding platform.
Fig. 4 is a schematic view of the assembly of a chip and a parallel pneumatic finger cylinder.
FIG. 5 is a schematic diagram of the connection between the multi-force field coupled cell tissue culture chip and the control system.
Detailed Description
The following detailed description is to be read in connection with the accompanying drawings and the detailed description:
example 1
The cell tissue culture chip for coupling the mechanical force field, the fluid force field and the hydrostatic force field is characterized in that the chip is fixed on a sliding platform of a parallel pneumatic finger cylinder through a screw and a three-hole chip clamp, the liquid inlet ends of the cylinder and the chip are connected with a gas control system, and the control system controls the on-off of an electromagnetic valve and the voltage application time to drive the chip to be stretched and pressurized.
Pouring the liquid PDMS prefabricated liquid (A glue: B glue is 10:1) with bubbles removed into a chip mold prepared in advance, placing on a horizontal table in a 65 ℃ blast oven, heating and curing for 90min, and taking out; taking out the chip from the mold, punching the positions of the inlet (10) and the outlet through hole (11) of the upper PDMS chip (2) shown in figure 1 by using a 2mm chip puncher, punching the positions of the screw through holes (9) and (8) of the upper PDMS chip (2) and the lower PDMS chip (1) shown in figure 1 by using a 3mm chip puncher, carrying out plasma treatment for 10min and then aligning and bonding under the condition that the bonding surfaces of the upper PDMS chip (2) and the lower PDMS chip (1) shown in figure 1 are clean, carrying out heat treatment in a 60 ℃ blast oven for 30min and then taking out, screwing a luer connector out, and completing the preparation of the chip after the liquid inlet through hole;
processing a 40mm × 15mm × 4mm acrylic cuboid on an acrylic plate with a thickness of 4mm by using a laser engraving machine, and forming a circular through hole with a d of 3.5mm on the cuboid, so as to obtain a three-hole chip clamp (as shown in fig. 2); the chip prepared above was fixed on the slide platform of the parallel pneumatic finger cylinder as shown in fig. 3 (1) using M3 stainless steel screws and three-hole chip clamp, and the assembly flow is shown in fig. 4; after the assembled system is subjected to ultraviolet treatment for 60min, injecting rat tail collagen diluted by 0.1% acetic acid into a round-corner rectangular cell tissue culture chamber from a luer connector screwed into a liquid inlet through hole by using an injector, and discharging redundant gas from the luer connector screwed into a liquid outlet through hole; after incubation for 30min at room temperature, sucking out collagen solution in the round-corner rectangular cell tissue culture chamber by using an injector, slowly injecting PBS into the round-corner rectangular cell tissue culture chamber from a luer connector screwed into the liquid inlet through hole by using the injector, and slowly sucking out the PBS; the operation is repeated for three times, so that the round-corner rectangular cell tissue culture chamber is ensured to have no residual acetic acid;
slowly injecting prepared cell tissue suspension into the round-corner rectangular cell tissue culture chamber from the luer connector screwed into the liquid inlet through hole, and blocking the luer connector screwed into the liquid outlet through hole by using a plug matched with the luer connector after gas in the chamber is discharged from the luer connector screwed into the liquid outlet through hole; the luer connector screwed into the liquid inlet through hole is connected with a hydraulic control system, the parallel pneumatic finger cylinder is connected with a gas control system (as shown in figure 5), and the parallel pneumatic finger cylinder assembled with the chip is placed in a carbon dioxide incubator at 37 ℃; and opening the switch of the control system, and controlling the switch of the electromagnetic valve and the voltage application time through the control system to drive the stretching of the chip and the increase of the liquid pressure in the chip, wherein the stretching amplitude and the liquid pressure can be modified and set in the control system according to the experimental requirements.
Experimental results show that the chip can realize a cell tissue culture scheme of independent application of mechanical force, hydrodynamics and hydrostatic pressure or coupling of multiple force fields, realize controllable reproduction of in-vitro cell tissue microenvironment, and can observe the response of cell tissues to parameter changes in the microenvironment in real time by combining the technologies of microscope observation, immunoassay and the like.
The foregoing is a description of features and embodiments of the present invention. The invention is not limited to the specific embodiments, but rather, variations and modifications are possible which fall within the scope of the invention as claimed.
Claims (9)
1. A multi-force field coupled cell tissue culture chip is characterized in that the multi-force field coupling is mechanical mechanics, hydromechanics and hydrostatic pressure multi-force field coupling;
the chip is composed of an upper-layer PDMS chip (2) and a lower-layer PDMS chip (1) which are mutually bonded, wherein the upper-layer PDMS chip (2) and the lower-layer PDMS chip (1) are rectangles with the same size; the upper binding surface of the lower PDMS chip (1) is provided with a round-corner rectangular cell tissue culture micro-cavity (4) with a downward opening, a liquid outlet cavity (5), a liquid inlet cavity (6), a liquid flow channel (7) and a lower chip screw through hole (8); an outlet (10), a liquid inlet through hole (11) and a screw through hole (9) which correspond to the groove flow channel of the lower-layer chip are formed in the upper-layer PDMS chip (2); the three-hole chip clamp (3) is a cuboid made of acrylic materials, and screw through holes (12) of the three-hole chip clamp are formed by using a laser engraving machine;
after the upper PDMS chip (2) and the lower PDMS chip (1) are bonded, the luer connector is screwed into the outlet (10) and the liquid inlet through hole (11), other reagents such as cell tissue suspension, cell tissue culture medium and the like are filled into the round-corner square cell tissue culture microcavity (4) from the luer connector screwed into the liquid inlet through hole (11), and redundant gas in the cavity is discharged through the luer connector screwed into the liquid outlet through hole (10); the luer connector screwed into the liquid through hole (11) is connected with a hydraulic control system; fixing the combined upper-layer (2) and lower-layer PDMS chips (1) on a parallel pneumatic finger cylinder sliding platform through a three-hole chip clamp (3) and a stainless steel screw; after the assembly is finished, the air cylinder is connected with the air pressure control system, the chip is connected with the hydraulic control system, and the control system controls the on-off of the electromagnetic valve and the voltage application time to drive the stretching movement of the chip and the internal pressurization of the chip.
2. The multi-force-field-coupled cell tissue culture chip according to claim 1, wherein the upper layer PDMS chip (2) is a cuboid with 88.5mm x 40mm x 4mm, the inlet (11) and outlet (10) through holes are 2mm through holes, and the screw through holes (9) are 3mm round through holes.
3. The multi-force-field-coupled cell tissue culture chip according to claim 1, wherein the lower PDMS chip (1) is a cuboid of 88.5mm × 40mm × 4mm, rectangular grooves with rounded corners (R ═ 1mm) of 22mm × 20mm × 1.7mm in cell tissue culture micro-cavities are formed downwards, the liquid flow channel (7) is formed by an arc with a depth of 1.7mm (R ═ 6.2 mm), the liquid inlet cavity (6) and the liquid outlet cavity (5) are circular with a depth of 1.7mm (R ═ 2 mm), the screw through hole (8) is a circular through hole with a depth of 1.7mm (d ═ 3mm, and the center of the screw through hole is aligned with the center of the screw through hole (9) of the upper PDMS chip (1).
4. The multi-force-field-coupled cell tissue culture chip according to claim 1, wherein the three-hole chip clamp (3) is a cuboid of 40mm x 15mm x 4mm acrylic, and the three-hole chip clamp (3) is provided with a circular through hole with d ═ 3.5mm by using a laser engraving machine, and the circle center of the circular through hole is aligned with the circle centers of the screw through holes (9), (8) of the upper-layer PDMS chip (2) and the lower-layer PDMS chip (1) and the threaded hole of the flat pneumatic finger cylinder.
5. The multi-force-field-coupled cell tissue culture chip of claim 1, wherein the bonding modes of the upper layer (2) and the lower layer PDMS chip (1) are plasma irreversible bonding.
6. The multi-force-field-coupled cell tissue culture chip of claim 1, wherein the upper layer (2) and the lower layer PDMS chip (1) are fixed on a parallel pneumatic finger cylinder sliding platform by using M3 stainless steel screws and a three-hole chip clamp (3).
7. The multi-force-field-coupled cell tissue culture chip according to claim 1, wherein the parallel pneumatic finger cylinder model is MHF-20D.
8. The multi-force-field coupled cell tissue culture chip of claim 1, wherein the parallel pneumatic finger cylinder is a gas-liquid combined cylinder, and realizes higher synchronization precision by the hydraulic incompressible characteristic.
9. The multi-force-field coupled cell tissue culture chip of claim 1, wherein the parallel pneumatic finger cylinder is a gas-liquid combined cylinder, and external mechanical connection is used to realize stable and reliable synchronous motion.
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| CN202010346127.4A CN113637580A (en) | 2020-04-27 | 2020-04-27 | Multi-force field coupled cell tissue culture chip |
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| CN202010346127.4A CN113637580A (en) | 2020-04-27 | 2020-04-27 | Multi-force field coupled cell tissue culture chip |
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| KR101887633B1 (en) * | 2017-03-09 | 2018-08-13 | 계명대학교 산학협력단 | Fluidic chip simulating wounds |
| CN208177479U (en) * | 2018-03-22 | 2018-12-04 | 广州安方生物科技有限公司 | A kind of microflow control chip clamping device |
| WO2019222871A1 (en) * | 2018-05-21 | 2019-11-28 | 深圳华大生命科学研究院 | Bionic intestinal-hepatic organ chip, preparation method therefor and application thereof |
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2020
- 2020-04-27 CN CN202010346127.4A patent/CN113637580A/en active Pending
Patent Citations (7)
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|---|---|---|---|---|
| CN102749443A (en) * | 2011-04-22 | 2012-10-24 | 国家纳米科学中心 | Double layer micro fluidic chip device and its application in immunodetection |
| CN103146576A (en) * | 2013-02-04 | 2013-06-12 | 中国科学院大学 | Micro-fluidic chip culture device of cell capable of being stretched mechanically and application thereof |
| CN105823727A (en) * | 2016-05-05 | 2016-08-03 | 北京纳迅科技股份有限公司 | Microfluid control detecting device and system |
| KR101887633B1 (en) * | 2017-03-09 | 2018-08-13 | 계명대학교 산학협력단 | Fluidic chip simulating wounds |
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