CN103146574B - A kind of high-throughput micro-fluidic biological mechanics long-time stimulus system and application thereof - Google Patents
A kind of high-throughput micro-fluidic biological mechanics long-time stimulus system and application thereof Download PDFInfo
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Abstract
The invention provides a kind of high-throughput micro-fluidic biological mechanics long-time stimulus system and application thereof, comprise at least one biomechanics and stimulate micro fluidic device, cell cultures drive system and negative pressure generator, described cell cultures drive system is connected with micro fluidic device with the liquid-flow driven in the microchannel of micro fluidic device; Described negative pressure generator is connected with microfluidic devices to make it produce negative pressure; And the high throughput testing of biomechanics stimulation study, related drugs and screening or for the preparation of the application in the test kit of biological detection in vitro.System of the present invention is based on microflow control technique, by the micro fluidic device of appropriate design and integrated micro flow channel module, elastica, negative pressure generation module, hydrodynamic shear and mechanical stretching force can be provided simultaneously, set up the external model of disease in body, for correlative study provides effective tool, be easy to make and use, multiple arteries simulation micro fluidic device can be connected simultaneously, realize high-throughput and observe and detect.
Description
Technical field
The present invention relates to a kind of high-throughput micro-fluidic biological mechanics long-time stimulus system and application thereof, belong to biomedicine technical field.
Background technology
Organism inner cell is in an environment being full of mechanical stimulation, biomechanics stimulates and take part in the normal physiological processes such as fetal development, osseous tissue balance, normal cardiovascular system held, and the biomechanics stimulation of exception then can cause generation, the development of arterial sclerosis, osteoporosis diseases.Although mechanical stimulation dysfunction is the important factor causing some disease, its mechanism of action it be unclear that, and the limitation of traditional research method hinders Developments.In recent years, the foundation of mechanical stimulation in vitro study model and application facilitate the progress of correlative study greatly.In body, modal two kinds of biomechanics stimulate is hydrodynamic shear and drawing force, nearly all vascular system as blood vessel, ureter and tissue space as all there are this two kinds of power in spatium interosseum etc.What current existing external model had can only Study of Fluid shearing force, as laminar flow plate, what also have can only study drawing force, but, be a complex environment having multiple mechanical stimulation in organism residing for cell, a biomechanics in vitro study model closer to internal milieu must consider the effect of multiple mechanical stimulation to cell.In recent years, people design and improve the device (Moore that some can apply hydrodynamic shear and Tensile stress simultaneously, J.E., Burki, E., Suciu, A., Zhao, S.M., Burnier, M., Brunner, H.R.and Meister, J.J. (1994) A Device for Subjecting Vascular Endothelial-Cells to Both FluidShear-Stress and Circumferential Cyclic Stretch.Ann Biomed Eng.22,416-422, Qiu, Y.C.and Tarbell, J.M. (2000) Interaction between wall shearstress and circumferential strain affects endothelial cell biochemicalproduction.J Vasc Res.37,147-157, Toda, M., Yamamoto, K., Shimizu, N., Obi, S., Kumagaya, S., Igarashi, T., Kamiya, A.and Ando, J. (2008) Differential gene responses in endothelial cells exposed to a combination ofshear stress and cyclic stretch.J Biotechnol.133, 239-244), its basic principle is the silicone rubber tube simulated blood vessel having attached endotheliocyte with inwall, keep applying Tensile stress by the expansion of tube chamber to the cell adhered to when certain pressure in tube chamber, apply shear-stress by washing away of liquid to cell simultaneously.But, the shortcoming of said apparatus also clearly, namely cell in the bad control of the adhesion of tube wall, Real Time Observation and intervention etc. can not be carried out to the behavior of cell under mechanical stimulation, these problems also govern the progress of correlative study.In recent years, the fast development of microflow control technique is that the foundation of numerous disease pathological study model provides opportunity, micro-fluidic chip can provide closer to the microenvironment under physiology, pathological conditions for cell, can mating surface chemistry and Soft lithograph technology cell behavior is manipulated and intervenes, observation and analysis can also be carried out at cell colony and unicellular two kinds of levels to the Behavioral change of cell.(Huh, D., the Matthews such as Huh, B.D., Mammoto, A., Montoya-Zavala, M., Hsin, H.Y.and Ingber, D.E. (2010) ReconstitutingOrgan-Level Lung Functions on a Chip.Science.328, micro fluidic device 1662-1668) adopted can produce hydrodynamic shear and drawing force to the cell be attached on film, and its purposes is simulation and research alveolar function.(the Douville such as Douville, N.J., Zamankhan, P., Tung, Y.C., Li, R., Vaughan, B.L., Tai, C.F., White, J., Christensen, P.J., Grotberg, J.B.and Takayama, S. (2011) Combination of fluid and solid mechanicalstresses contribute to cell death and detachment in a microfluidic alveolarmodel.Lab Chip.11, micro fluidic device 609-619) adopted can provide hydrodynamic shear and mechanical stretching force equally, its purposes is also the structure and function of simulation and research alveolar.But above-mentioned micro fluidic device is only suitable for making alveolar model, and be difficult to carry out long-time experiment cell being carried out to mechanical stimulation.Because long-time micro-current controlled cell cultivates and experiment faces the problems such as the control of such as temperature and potential of hydrogen, bubble generation, blocking, leakage, wherein any one problem all may cause the failure of experiment.
Summary of the invention
Therefore, the object of the invention is to be only suitable for making alveolar model for existing micro fluidic device, and be difficult to carry out long-time deficiency of cell being carried out to the experiment of mechanical stimulation, there is provided one to provide hydrodynamic shear and drawing force simultaneously, and the microfluidic system that long-time biomechanics can be provided to stimulate and application thereof, for correlative study provides effective tool.
For above-mentioned technical purpose, technical scheme of the present invention is as follows:
On the one hand, the invention provides a kind of high-throughput micro-fluidic biological mechanics long-time stimulus system, comprise at least one biomechanics and stimulate micro fluidic device, cell cultures drive system and negative pressure generator, described cell cultures drive system is connected with micro fluidic device with the liquid-flow driven in the microchannel of micro fluidic device; Described negative pressure generator is connected with microfluidic devices to make it produce negative pressure, and the physiology of its organ stimulated for biomechanics, tissue, the high-throughput of pathological condition are observed and detected.
Preferably, described biomechanics stimulates micro fluidic device to comprise transparent microchannel module and the transparent negative pressure generation module suitable with it, described microchannel module bottom and negative pressure generation module top are by elastica phase bonding, described biomechanics stimulation micro fluidic device is the intasome of described microchannel module, described elastica, described negative pressure generation module, described microchannel module is used for fluid flowing, and described negative pressure generation module is for generation of the negative pressure making elastica generation deformation;
Described microchannel module top is provided with fluid outlet and fluid intake, described fluid outlet and fluid intake are connected to microchannel respectively by with fluid intake and the suitable PE pipe of fluid outlet, and connect with it, described microchannel is located at described microchannel module bottom;
Described negative pressure generation module top is provided with negative pressure groove, and described negative pressure groove is located at below microchannel, and described negative pressure generation module top is provided with a gas circuit opening, and described gas circuit opening is by being connected with negative pressure groove with its 2nd suitable PE pipe.
Preferably, described cell cultures drive system comprises liquid storage bottle and connected peristaltic pump, and described biomechanics stimulates micro fluidic device, described liquid storage bottle, described peristaltic pump to form circulating fluid path by a described PE pipe series connection; Described liquid storage bottle is connected with described microchannel with described fluid outlet by a described PE pipe, described peristaltic pump is connected with described liquid storage bottle by a described PE pipe, and described peristaltic pump is connected to drive the fluid in microchannel to flow with described microchannel with described fluid intake by a PE pipe.
Preferably, described negative pressure generator comprises air-flow distributor, gas flow controller and minipump, is connected to gas flow opening and negative pressure generation model calling bottom described air-flow distributor by the 2nd PE pipe, produces negative pressure to make negative pressure generation module; Described gas flow controller comprises controller and driven airflow switch, and described air-flow distributor top and minipump are all connected to the first end of driven airflow switch, and the second end of described driven airflow switch is connected with controller.
Preferably, described negative pressure groove side is also provided with the negative pressure Buffer Pool connected with it, and described 2nd PE pipe is connected with negative pressure groove by negative pressure Buffer Pool.
Preferably, described negative pressure Buffer Pool is connected by a groove and negative pressure groove, and preferably, described 2nd PE pipe is vertically connected at bottom negative pressure Buffer Pool.
Preferably, around a rectangular platform in the middle of described negative pressure groove, and described elastica is close to and corresponds to the described microchannel above it, is filled with liquid lubricant between described rectangular platform and described elastica; Preferably, the top of described negative pressure generation module and bottom involution and into a single integrated structure, described rectangular platform length is 1.5 × 10
4μm, wide is 1.0 × 10
3μm.
Preferably, described microchannel is the second rectangular structure, and it, by bottom the 3rd, the 4th sidewall and microchannel, is made up of elastica bottom described microchannel, and preferably, the length of described second rectangular parallelepiped is 1.8 × 10
4μm, wide is 1.5 × 10
3μm, height is 0.5 × 10
3μm.
Preferably, the top of described microchannel module and bottom connect into the first through rectangular structure of horizontal direction by first, second sidewall, first, second sidewall described and top are by polydimethylsiloxane (PDMS) module composition, preferably, the length of described first rectangular parallelepiped is 2.5-3.0 × 10
4μm, wide 2.0-2.5 × 10
4μm, height is 3.0-5.0 × 10
3μm.
Preferably, described microchannel module, negative pressure generation module and elastica are made by polydimethylsiloxane (PDMS) material.
Preferably, described fluid outlet and fluid intake are circular port, and preferably, the diameter of described circular port is 8.0 × 10
2μm; A described PE pipe is vertically connected at described microchannel module bottom.
Preferably, the top of described negative pressure generation module is rectangle, and this rectangular length is 2.5-3.0 × 10
4μm, wide is 2.0-2.5 × 10
4μm, described negative pressure groove is the 3rd rectangular structure, and its cross section is rectangle, and height is 5 × 10
2μm, wide is 2.5 × 10
2μm; Described negative pressure Buffer Pool is in the cylindrical cavity shape perpendicular to platform area, and the diameter of described cylindrical cavity is 5.0 × 10
3μm, height is 1.0 × 10
3μm; Described groove is the 4th rectangular structure, and its length is 3.0 × 10
3μm, wide is 5 × 10
2μm, height is 5 × 10
2μm; Described gas circuit opening is circular port, and preferably, the diameter of described circular port is 8 × 10
2μm.
Preferably, described microchannel film block and negative pressure produce film block length and are 2.5-3.0 × 10
4μm, be widely 2.0-2.5 × 10
4μm, be thickly 3.0-5.0 × 10
3μm; Described elastica is long is 2.5-3.0 × 10
4μm, wide 2.0-2.5 × 10
4μm, thick is 10-100 μm.
On the other hand, the invention provides a kind of high-throughput micro-fluidic biological mechanics long-time stimulus system in the high throughput testing of biomechanics stimulation study, related drugs and screening or for the preparation of the application in the test kit of biological detection.
Another aspect, the invention provides a kind of micro-fluidic biological mechanical stimulation system for the preparation of the application in the test kit of biological detection, preferably, described test kit is be the research of external biological mechanical stimulation or the high throughput testing of related drugs and the test kit of screening.
Again on the one hand, the invention provides a kind of test kit for biological detection, this test kit comprises according to high-throughput micro-fluidic biological mechanics long-time stimulus system of the present invention, also comprise detection reagent and damping fluid, preferably, described detection reagent is cytokine, antibody, active small molecular or the medicine for screening that cyto-mechanics is correlated with.
Beneficial effect of the present invention is: based on microflow control technique, micro fluidic device is stimulated by the biomechanics of appropriate design and integrated micro flow channel module, elastica, negative pressure generation module, hydrodynamic shear and mechanical stretching force can be provided simultaneously, a system can comprise multiple micro fluidic device simultaneously, the high-throughput of the organ stimulated for biomechanics, the physiology of tissue, pathological condition is observed and is detected, set up the external model of disease in body, for correlative study provides effective tool, volume is little, structure is simple, is easy to make and use; Optically transparent material makes, be easy to the situation in naked eyes or Microscopic observation passage, can change arbitrarily between microscope and incubator, can realize the original position dynamic monitoring of cell under two kinds of mechanical stimulations and under chemical stimulation, size, the frequency of two kinds of mechanical stimulations are adjustable at any time, can change the chemical micro-environment of cell at any time, temperature, acid or alkali environment are constant, and system can keep long-time stable work; Realize high-throughput to observe and detect.
Accompanying drawing explanation
Below, describe embodiment of the present invention in detail by reference to the accompanying drawings, wherein:
Fig. 1 is the structural representation of high-throughput micro-fluidic biological mechanics long-time stimulus system of the present invention;
Fig. 2 is the structural representation that biomechanics of the present invention stimulates micro fluidic device;
Fig. 3 is that described elastica is positioned in the micro fluidic device of system of the present invention the experimental result schematic diagram carrying out stretching, and in figure, a is the result schematic diagram of the elastica before stretching, and in figure, b is the experimental result schematic diagram of the elastica after stretching;
Fig. 4 to stretch separately with MSC cell after in the micro-fluidic biological mechanical stimulation device of system of the present invention for be inputted by Bone Marrow Stromal Stem Cells (MSC cell) and to shear separately the experimental result schematic diagram carrying out contrasting, in figure, a is the experimental result schematic diagram that MSC cell stretches separately, b is the experimental result schematic diagram that MSC cell is sheared separately, and c is MSC cell drawn and the experimental result schematic diagram after shearing in the apparatus of the present;
Wherein:
1 is microchannel module, 101 is the top of microchannel module, 102 is the bottom of microchannel module, 103 is the first side wall, and 104 is the second sidewall, and 105 is fluid intake, 106 is fluid outlet, 107 is a PE pipe, and 108 be microchannel, 1081 is microchannel top, 1082 is bottom microchannel, 1083 be the 3rd sidewall, 1084 is the 4th sidewall;
2 is elastica;
3 is negative pressure generation module, and 301 is the bottom of negative pressure generation module, and 302 is the top of negative pressure generation module, and 303 is negative pressure groove, and 304 is gas flow opening, 303 negative pressure grooves, and 305 is the 2nd PE pipe, 306 negative pressure cushion chambers, and 307 is groove, and 308 is rectangular platform;
4 is biomechanics stimulation micro fluidic device; 5 is liquid storage bottle; 6 is peristaltic pump; 7 is air-flow distributor; 8 is controller; 9 is driven airflow switch; 10 is minipump.
Embodiment
As shown in Figure 1, high-throughput micro-fluidic biological mechanics long-time stimulus system of the present invention, comprise cell cultures drive system 5, negative pressure generator 6 and biomechanics and stimulate micro fluidic device 4, described cell cultures drive system and biomechanics stimulate micro fluidic device to be connected to drive biomechanics to stimulate liquid-flow in the microchannel of micro fluidic device; Described negative pressure generator and biomechanics stimulate microfluidic devices to be connected to make it produce negative pressure.
Described cell cultures drive system comprises liquid storage bottle 5 and connected peristaltic pump 6, described liquid storage bottle 5 is connected with microchannel 108 by fluid intake 105, and described peristaltic pump 6 is connected to drive the fluid in microchannel to flow by fluid outlet 106 with microchannel 108.
Described negative pressure generator comprises air-flow distributor 7, gas flow controller and minipump 10, is connected to gas flow opening 304 is connected with negative pressure generation module 3 bottom described air-flow distributor 7 by the 2nd PE pipe 305, produces negative pressure to make negative pressure generation module 3; Described gas flow controller comprises controller 8 and driven airflow switch 9, and described air-flow distributor 7 top and minipump 10 are all connected to the first end of driven airflow switch 9, and the second end of described driven airflow switch 9 is connected with controller 8.
The capacity of described liquid storage bottle 501 is 50 milliliters, and bottle cap and bottle can tighten sealing, and bottle cap has three holes, is respectively liquid flow hand-hole, liquid outflow port, air-vent.Liquid inflow port and taphole are 8.0 × 10 by diameter
2μm PE pipe through, two Guan Jun are opened on below liquid level, by cast PDMS and sealing between PE pipe with bottle cap.Air-vent is 8.0 × 10 by internal diameter
2μm syringe needle pass, syringe needle part in bottle is positioned at more than liquid level, and the other end and the aperture of syringe needle are that the pin type filter of 0.22 micron is connected, and are opened in air.Syringe needle passes covering part and syringe needle and filter connection portion and all seals.
The specification of described peristaltic pump 502: minimum flow rate can be provided to be 2 mul/min, and roller bearing is 10, two passages.
Described minipump 603, flow is 28 liters/min, and vacuum tightness is 10 kPas.
The similar syringe of described driven airflow switch 6022, is made up of cylindrical shell and piston.Sheating material is copper, and internal diameter is 1 centimetre, and long 5 centimetres, have two openings in bottom, internal diameter is 5 millimeters, connects the two ends near, far away of extraction pipe respectively.There is an opening at the middle part (2.5 centimeters) of shell, and internal diameter is 3 millimeters.Piston portion material is stainless steel, and length is 6 centimetres, and external diameter is 0.5 centimetre, and piston crown material is tetrafluoroethylene, and the internal diameter of its external diameter and shell is suitable, and between housing, tight friction forms the sealing to gas.The afterbody of piston is connected with the disk be fixed on step motor shaft by bearing and connecting rod.Stepper-motor is connected by signal wire with controller 6021, and controller 6021 can regulate motor speed and direction.
Described air-flow distributor: made by PDMS material, volume is external diameter is 2 centimetres, and height is the right cylinder of 2 centimetres, and interior is diameter 1 centimetre, and height is the cylindrical cavity of 1 centimetre.Directly over cylindrical, have a hole, connect the PE pipe that internal diameter is 2 millimeters, this pipe is connected to the distal openings of gas flow controller by aperture conversion joint.On the cylindrical surface of cylindrical cavity, open 4-6 hole respectively and be connected with the PE pipe of 0.8 mm outer diameter, 0.5 millimeter of internal diameter, this PE pipe is connected to the negative pressure air vent openings of micro flow chip, and multiple PE pipe can connect multiple chip.
As shown in Figure 2, micro fluidic device 4 of the present invention, this device comprises transparent microchannel module 1 and the transparent negative pressure generation module 3 suitable with it, elastica 2 is passed through through plasma oxidation process covalent bonding in described microchannel module bottom and negative pressure generation module top, described microchannel module 2, negative pressure generation module 3 and elastica 2 are made by PDMS material, and described springform 2 is long 2.5-3.0 × 10
4μm, wide 2.0-2.5 × 10
4μm, thick is the springform 2 of 10-100 μm, described microchannel module 1 is for fluid flowing, and described negative pressure generation module 3 is for generation of the negative pressure making elastica 2 that deformation occur, described microchannel module top 101 and bottom 102 connect into the first through rectangular structure of horizontal direction by first, second sidewall 103,104, and the length of described first rectangular parallelepiped is 2.5-3.0 × 10
4μm, wide 2.0-2.5 × 10
4μm, height is 3.0-5.0 × 10
3μm, described microchannel module is long 2.5-3.0 × 10
4μm, wide 2.0-2.5 × 10
4μm, thick is 3.0-5.0 × 10
3μm PDMS module, described microchannel module top 101 is provided with fluid intake 105 and fluid outlet 106, described fluid intake 105 and fluid outlet 106 are vertically connected at the microchannel 108 of microchannel module bottom 102 respectively by with fluid intake 105 and the suitable PE pipe 107 of fluid outlet 106, described microchannel 108 is in the second rectangular structure, it is by microchannel top, bottom 1081, 1082 and the 3rd, 4th sidewall 1083, 1084 are formed, and connect with it, be made up of elastica bottom described microchannel, the length of described second rectangular parallelepiped is 1.8 × 10
4μm, wide is 1.5 × 10
3μm, height is 0.5 × 10
3μm, described fluid intake 105 and fluid outlet 106 are circular port, and the diameter of described circular port is 8.0 × 10
2μm, the bottom 301 of described negative pressure generation module 3 and top 302 involution and into a single integrated structure, described negative pressure generation module 3 is by PDMS module composition, and described negative pressure generation module is long is 2.5-3.0 × 10
4μm, wide is 2.0-2.5 × 10
4μm, thick is 3.0-5.0 × 10
3μm, described negative pressure generation module top 302 is provided with negative pressure groove 303, described negative pressure groove 303 is located at the below of microchannel 108, thus form the independent space isolated with microchannel 108, described negative pressure generation module top 302 is provided with a gas circuit opening 304, described gas circuit opening 304 is by being connected with negative pressure groove 303 with its 2nd suitable PE pipe 305, around a rectangular platform 308 in the middle of described negative pressure groove, be close to described elastica and correspond to the described microchannel 108 above it, rectangular platform length is 1.5 × 10
4μm, wide is 1.0 × 10
3μm, described rectangular platform 308 and elastica 2 not bonding, adding liquid lubricant, make it can move each other under suction function, the negative pressure Buffer Pool 306 connected with it is also located in described negative pressure groove 303 side, described 2nd PE pipe 305 is connected with negative pressure groove 303 by negative pressure Buffer Pool 306, described negative pressure Buffer Pool 306 is connected by a groove 307 and negative pressure groove 303, described 2nd PE pipe 305 is vertically connected at bottom negative pressure Buffer Pool 306, , the top 302 of described negative pressure generation module is rectangle, this rectangular length is 2.5-3.0 × 10
4μm, wide is 2.0-2.5 × 10
4μm, described negative pressure groove 303 is in the 3rd rectangular structure, and its cross section is rectangle, and height is 5.0 × 10
2μm, wide is 2.5 × 10
2μm, described negative pressure Buffer Pool 306 is in the cylindrical cavity shape perpendicular to bottom 301, and the diameter of described cylindrical cavity is 5.0 × 10
3μm, height is 1.0 × 10
3μm, described groove 307 is in the 4th rectangular structure, and its length is 3.0 × 10
3μm, wide is 5.0 × 10
2μm, height is 5.0 × 10
2μm, described gas circuit opening 304 is circular port gas circuit opening, and the diameter of described circular port gas circuit opening 304 is 8.0 × 10
2μm.
During use, first by the fluid intake 105 of microchannel module 1, cell suspension is added microchannel 108, under 37 degree Celsius of 5% carbon dioxide conditions, make in the elastica 2 of cell attachment bottom microchannel 108, then, the fluid intake 105 of microchannel 108 is connected with cell culture medium drive system 5 with fluid outlet 106, again the gas circuit opening 304 of negative pressure generation module 3 is connected with negative pressure generator 6 by the 2nd PE pipe, then, start cell culture medium drive system 5, cell culture medium drive system 5 can drive the liquid-flow in microchannel 108, thus hydrodynamic shear is produced to the cell that substrate in microchannel 108 is adhered to, and the negative pressure that negative pressure generator 6 produces can be conducted in the negative pressure groove 303 in negative pressure generation module 3 by the 2nd PE pipe 305, make the elastica 2 above negative pressure groove 303 that deformation occur, thus pull the elastica 2 being affixed on rectangular platform, make it the deformation in occurred level direction, thus make the superincumbent cell of attaching be subject to mechanical stretching force.Because elastica 2 is to be parallel to the deformation on passage long axis direction very little, negligible, if what be applied to cell in elastica 2 and elastica advocates perpendicular to passage long axis direction, therefore, the stressing conditions of cell is: be parallel to the hydrodynamic shear of channel direction and the mechanical stretching force perpendicular to channel direction.
concrete test example
test example 1
By elastica (PDMS film, fluorescent marker protein dot matrix is printed on above it) be positioned in the micro fluidic device of high-throughput micro-fluidic biological mechanics long-time stimulus system of the present invention and stretch, result as shown in Figure 3, PDMS film after negative pressure stretches, compared with the PDMS film before stretching, extensibility reaches 25%, and its extensibility and homogeneity can produce a desired effect, and meets the needs of simulation in vivo test completely.
test example 2
By MSC cell (Bone Marrow Stromal Stem Cells, take from SD rat (purchased from Beijing laboratory animal company of dimension tonneau China), extracting method is with reference to carrying out with Publication about Document: Bosnakovski D, Mizuno M, Kim G, Takagi S, Okumura M, Fujinaga T.Isolation and multilineagedifferentiation of bovine bone marrow mesenchymal stem cells.Cell Tissue Res2005; 319:243-53.) input in the micro fluidic device of high-throughput micro-fluidic biological mechanics long-time stimulus system of the present invention, stretch separately with MSC cell and to compare with the result sheared separately, as shown in Figure 4, stretch separately as can be seen from this figure and the skeleton of cell can be made along the arrangement of drawing force parallel direction, and hydrodynamic shear can make cytoskeleton arrange along shearing force direction separately; Drawing force then can make the arrangement of cytoskeleton present the trend identical with resultant direction with hydrodynamic shear with joint efforts.Therefore, hydrodynamic shear and drawing force are arranged with material impact to intravascular cells, thus illustrate that system of the present invention is the powerful of correlative study.
Claims (20)
1. a high-throughput micro-fluidic biological mechanics long-time stimulus device, it is characterized in that, comprise cell cultures drive unit, negative pressure generator and at least one biomechanics and stimulate micro fluidic device, described cell cultures drive unit and biomechanics stimulate micro fluidic device to be connected with the liquid-flow driven in the microchannel of micro fluidic device; Described negative pressure generator and biomechanics stimulate micro fluidic device to be connected to make it produce negative pressure
Wherein, described biomechanics stimulates micro fluidic device to comprise transparent microchannel module and the transparent negative pressure generation module suitable with it, described microchannel module bottom and negative pressure generation module top are by elastica phase bonding, described microchannel module is used for fluid flowing, and described negative pressure generation module is for generation of the negative pressure making elastica generation deformation;
Wherein, around a rectangular platform in the middle of described negative pressure groove, be close to described elastica and correspond to the described microchannel above it, between described rectangular platform and elastica, being filled with liquid lubricant;
Described microchannel is the second rectangular structure, and it is formed by bottom the 3rd, the 4th sidewall and microchannel, is made up of bottom described microchannel elastica;
Described microchannel module top is provided with fluid outlet and fluid intake, described fluid outlet and fluid intake are connected to microchannel respectively by with fluid intake and the suitable PE pipe of fluid outlet, and connect with it, described microchannel is located at described microchannel module bottom;
Described negative pressure generation module top is provided with negative pressure groove, and described negative pressure groove is located at below microchannel, and described negative pressure generation module top is provided with a gas circuit opening, and described gas circuit opening is by being connected with negative pressure groove with its 2nd suitable PE pipe.
2. device according to claim 1, it is characterized in that, described cell cultures drive unit comprises liquid storage bottle and connected peristaltic pump, described liquid storage bottle, described peristaltic pump form circulating fluid path by a described PE pipe series connection, described liquid storage bottle is connected with described microchannel with described fluid outlet by a described PE pipe, described peristaltic pump is connected with described liquid storage bottle by a described PE pipe, and described peristaltic pump is connected to drive the fluid in described microchannel to flow with described microchannel with described fluid intake by a PE pipe.
3. device according to claim 1, it is characterized in that, described negative pressure generator comprises air-flow distributor, gas flow controller and minipump, is connected to gas flow opening and negative pressure generation model calling bottom described air-flow distributor by the 2nd PE pipe, produces negative pressure to make negative pressure generation module; Described gas flow controller comprises controller and driven airflow switch, and described air-flow distributor top and minipump are all connected to the first end of driven airflow switch, and the second end of described driven airflow switch is connected with controller.
4. device according to claim 1, is characterized in that, described negative pressure groove side is also provided with the negative pressure Buffer Pool connected with it, and described 2nd PE pipe is connected with negative pressure groove by negative pressure Buffer Pool.
5. device according to claim 4, is characterized in that, described negative pressure Buffer Pool is connected by a groove and negative pressure groove.
6. device according to claim 4, is characterized in that, described 2nd PE pipe is vertically connected at bottom negative pressure Buffer Pool.
7. device according to claim 1, is characterized in that, the top of described negative pressure generation module and bottom involution and into a single integrated structure, and described rectangular platform length is 1.5 × 10
4μm, wide is 1.0 × 10
3μm.
8. device according to claim 1, is characterized in that, the top of described microchannel module and bottom connect into the first through rectangular structure of horizontal direction by first, second sidewall.
9. device according to claim 8, is characterized in that, the length of described first rectangular parallelepiped is 2.5-3.0 × 10
4μm, wide 2.0-2.5 × 10
4μm, height is 3.0 ~ 5.0 × 10
3μm.
10. device according to claim 1, is characterized in that, the length of described second rectangular parallelepiped is 1.8 × 10
4μm, wide is 1.5 × 10
3μm, height is 0.5 × 10
3μm.
11. devices according to any one of claim 1 to 10, is characterized in that, described microchannel module, negative pressure generation module and elastica are made by PDMS material.
12. devices according to any one of claim 1 to 10, is characterized in that, described fluid outlet and fluid intake are circular port.
13. devices according to claim 12, is characterized in that, the diameter of described circular port is 8.0 × 10
2μm; A described PE pipe is vertically connected at described microchannel module bottom.
14. devices according to any one of claim 1 to 10, is characterized in that, the top of described negative pressure generation module is rectangle, and this rectangular length is 2.5-3.0 × 10
4μm, wide is 2.0-2.5 × 10
4μm, described negative pressure groove is the 3rd rectangular structure, and its cross section is rectangle, and height is 5.0 × 10
2μm, wide is 2.5 × 10
2μm; Described negative pressure Buffer Pool is in the cylindrical cavity shape perpendicular to platform area, and the diameter of described cylindrical cavity is 5.0 × 10
3μm, height is 1.0 × 10
3μm; Described groove is the 4th rectangular structure, and its length is 3.0 × 10
3μm, wide is 5.0 × 10
2μm, height is 5.0 × 10
2μm; Described gas circuit opening is circular port.
15. devices according to claim 14, is characterized in that, the diameter of described circular port is 8.0 × 10
2μm.
16. devices according to any one of claim 1 to 10, is characterized in that, described microchannel module and negative pressure generation module length are 2.5-3.0 × 10
4μm, be widely 2.0-2.5 × 10
4μm, height is 3.0-5.0 × 10
3μm; Described elastica is long is 2.5-3.0 × 10
4μm, wide is 2.0-2.5 × 10
4μm, thick is 10-100 μm.
17. devices according to any one of claim 1 to 16 are studied at the external biological mechanical stimulation of non-diagnostic object, the high throughput testing of related drugs and screening or for the preparation of the test kit of biological detection in apply.
18. application according to claim 17, is characterized in that, described test kit is the research of external biological mechanical stimulation and the high throughput testing of related drugs and the test kit of screening.
19. 1 kinds, for the test kit of biological detection, is characterized in that, described test kit comprises the high-throughput micro-fluidic biological mechanics long-time stimulus device according to any one of claim 1 to 16, also comprises detection reagent and damping fluid.
20. test kits according to claim 19, is characterized in that, described detection reagent is cytokine, antibody, active small molecular or the medicine for screening that cyto-mechanics is correlated with.
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| CN103805511B (en) * | 2014-02-18 | 2015-10-28 | 国家纳米科学中心 | The arteries simulation micro fluidic device directly can observed under high power objective |
| CN105974102A (en) * | 2016-04-28 | 2016-09-28 | 新乡医学院 | Apparatus for observing shear stress adjusted PCSK9 expression |
| CN106554902B (en) * | 2016-09-02 | 2019-03-05 | 四川大学 | A kind of power-chemical Coupling biology experimental installation |
| CN108485973A (en) * | 2018-06-20 | 2018-09-04 | 上海长海医院 | A kind of cell co-cultures flat flow chamber and its fluid shear stress stimulating method |
| CN110438004B (en) * | 2019-09-04 | 2024-07-30 | 北京航空航天大学 | Flat plate flow cavity for simultaneously applying static traction force and flow shearing force |
| CN111426821B (en) * | 2020-03-24 | 2021-07-06 | 大连理工大学 | Micro-fluidic chip-level extracorporeal circulation system for vascular endothelial cell mechanics biology research |
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