CN102936567A - Microtopological-structure plate flow chamber capable of applying electric and shearing force stimulation - Google Patents

Microtopological-structure plate flow chamber capable of applying electric and shearing force stimulation Download PDF

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Publication number
CN102936567A
CN102936567A CN2012104179030A CN201210417903A CN102936567A CN 102936567 A CN102936567 A CN 102936567A CN 2012104179030 A CN2012104179030 A CN 2012104179030A CN 201210417903 A CN201210417903 A CN 201210417903A CN 102936567 A CN102936567 A CN 102936567A
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shearing force
topological framework
battery lead
flow chamber
lead plate
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CN102936567B (en
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吕永钢
董单娟
邹杨
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Chongqing University
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Chongqing University
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS 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
    • C12M35/00Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/02Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS 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
    • C12M21/00Bioreactors or fermenters specially adapted for specific uses
    • C12M21/08Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS 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
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/02Form or structure of the vessel
    • C12M23/04Flat or tray type, drawers
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS 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
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/02Form or structure of the vessel
    • C12M23/12Well or multiwell plates
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS 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
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/20Material Coatings

Abstract

The invention discloses a microtopological-structure plate flow chamber capable of applying electric and shearing force stimulation, which is formed by superposing three plates and two gaskets, wherein an upper plate (2) is provided with a fluid inlet/outlet port; an upper gasket (3) is provided with a diversion trench (4); a middle plate (5) is provided with a rectangular slit (6); a lower gasket (10) is provided with a flow cavity (18); the middle plate (5) above the flow cavity (18) is provided with a plate electrode (7); a lower plate (11) below the flow cavity (18) is provided with a plate electrode (16); a pair of electrodes (14 and 17) are respectively arranged on four sides of the flow cavity (18); and the plate electrode (16) is composed of a sheet (22), a PDMS (polydimethylsiloxane) film (23) and a graphene film (24), and has a microtopological structure on the surface. When applying electric and shearing force stimulation with different modes and intensities, the invention implements microtopological structure regulation on the cell (19). The structure of the device is simple, compact and reliable.

Description

Can apply the flat flow chamber with little topological framework that electricity and shearing force stimulate
Technical field
The present invention relates to a kind of flat flow chamber, particularly can apply the flat flow chamber with little topological framework that electricity and shearing force stimulate.
Background technology
Organizational project is the principle according to cytobiology and engineering science, in conjunction with seed cell, biologic bracket material and somatomedin at external/construct in vitro tissue and organ, to keep, repair, to regenerate or to improve damaged tissue and organ dysfunction.The biologic bracket material of compound seed cell is the core of organizational project, but numerous researchs have shown all biological functions such as adhesion, propagation and differentiation of regulating cell such as the little topological framework of matrix, electricity irritation and shearing force, can promote the interaction of seed cell and timbering material, the secretion of extracellular matrix in the regulating cell.In organizational project, usually utilize at present a kind of device can only realize a kind of stimulation mode.Flat flow chamber is the external main device that cell is applied the hydrodynamic shear stimulation.For cell is applied electricity irritation, can be in flow cavity electrode insertion, the electro-conductive material such as conductive glass etc. be installed realized.
But cell is to be subject to simultaneously the little topological framework of matrix, electricity irritation and shear action in a lot of situations in organizer.Take bone tissue restoration as example, the intercellular washing fluids that bone matrix deformation the causes formed hydrodynamic shear that flows is the main stress that cell can be experienced, and can affect the propagation, differentiation, mineralising of multiple seed cell etc.Ubiquity endogenous physiological electric field in the biological tissue organ, ectogenic electrical field stimulation also are used for promoting seed cell propagation, differentiation and with the adhesion of timbering material etc.The topological framework on timbering material surface can change the stress distribution of cell surface, thereby changes the form of cell, affects the alignment capabilities of propagation, differentiation, orientation, migration and the cytoskeleton of cell.Meanwhile, the response that cell stimulates different factors is not merely the linear superposition of single factors impact, can not only carry out the stimulation of single factors with single device in the research, need to study simultaneously the synergy of many factors.Therefore, urgent hope has a kind of device can realize simultaneously that many factors stimulates, with the response of conveniently observing and the research cell stimulates simultaneously to many factors.Yet yet there are no flow cavity and when cell being applied the stimulation of electricity and shearing force, realize little topological framework regulation and control.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of flat flow chamber with little topological framework that simultaneously cell is applied electricity and hydrodynamic shear stimulation, not only can apply a plurality of direction electricity and hydrodynamic shear stimulates, and can apply little topological framework stimulation to cell.
For solving the problems of the technologies described above, the flat flow chamber with little topological framework that applies electricity and shearing force stimulation of the present invention comprises three layer flat plate and two-layer pad, upper flat plate is provided with fluid inlet and outlet, Upper gasket is provided with diversion trench, middle plateform arranges rectangular slot, and lower gasket is provided with flow cell.Nutrient solution enters Upper gasket intermediate cam shape diversion trench by the upper flat plate fluid inlet, so that fluid is fully dispersed and the effect that cushioned effectively reduces the flow-disturbing phenomenon of entrance.The nutrient solution of fully dispersing enters the fluid cell by rectangular slot on the middle plateform.Satisfy laminar flow, two-dimensional flow, flowing of fully dispersing when guaranteeing the moving cell of fluid flows, flow cell width and length all should and be placed on thickness poor of battery lead plate wherein greater than flow cell thickness.For further realizing multidirectional electricity irritation, described middle plateform is provided with battery lead plate above being positioned at flow cell, and the placement below being positioned at flow cell of lower flat board has the battery lead plate of little topological framework, and an electrode is placed respectively on flow cell four limits.
The battery lead plate of placing on the lower flat board is by being fixed in thin slice and having the dimethyl siloxane (Polydimethylsiloxane of little topological framework, PDMS) table lining graphene film obtains on the film, the graphene film two ends arrange respectively the connection wire, can load electricity irritation and apply simultaneously little topological framework stimulation, wherein thin slice is made by silicon chip or slide.This electrode can be prepared into the flat board of consistency of thickness, realizes the hydrodynamic shear effect of homogeneous; Also can be prepared into the clapboard that even thickness changes, realize continuous shear action.Little topological framework can be micro-cannelure array as required, also can be the micro-boss array of trilateral, tetragon, pentagon, hexagon or all kinds of shapes such as cylindrical.When carrying out the flow cavity experiment, at graphene film surface seeding cell, perhaps as required lining is shown on the graphene film surface and inoculated cell.The middle plateform electric pole plate is by being adhesive on the middle plateform, and it connects wire draws by the sulculus that is engraved on the middle plateform, utilizes sealant sealing.Battery lead plate all can be made by graphene film or indium tin oxide conductive glass or indium tin oxide conductive film or the noble metal plate such as silver, gold or platinum around middle plateform electric pole plate and the flow cell.Connect that the wire of battery lead plate is laid on the sulculus of carving on the lower gasket around the flow cell, use sealant sealing after placing wire.
After flat flow chamber is good by bolt or Fixture assembly, by the system that circulates of one of teflon catheter and reservoir and peristaltic pump formation.After liquid in the reservoir is pumped by peristaltic pump, flow into flat flow chamber along conduit, the cell on the battery lead plate is applied shear action, then flow out, get back in the reservoir along flowing out conduit.Shearing force loading mode (continuously, vibration or pulse etc.) and intensity are controlled by peristaltic pump.During work, except peristaltic pump, other devices all are placed in the incubator.By signal generator and voltage stabilized source give each to electrode or unitary electrode apply continuously, the electricity irritation of the multiple binding mode such as vibration or pulse and intensity.Can only use an electrode during electricity irritation, also can use pair of electrodes, can also use simultaneously a plurality of electrodes.
In sum, flat flow chamber of the present invention passes through at six face installing electrodes of flow cell plate, and the lower dull and stereotyped upper battery lead plate of placing has little topological framework, cell is realized that simultaneously electricity and shear action and little topological framework stimulate, can also realize homogeneous hydrodynamic shear or continuous shear action by changing the lower dull and stereotyped upper battery lead plate thickness of placing, this also is one of key of proposing of the present invention.Flat flow chamber provided by the invention can be selected different little topological frameworks according to demand, applying simultaneously different mode and intensity electricity and hydrodynamic shear stimulates, the flat flow chamber of realizing is simple and compact for structure, is expected to play a significant role in more Tissue Engineering Study.Material for the preparation of the electrode that is placed on the flow cell bottom not only is confined to graphene film, as long as the material of selecting can form little topological framework and conduction, cell is got final product without overt toxicity.For for purpose of brevity, the present invention is mainly set forth as an example of graphene film example, and other electro-conductive materials also can adopt identical principle.
Description of drawings
Fig. 1 is the structural representation that applies the flat flow chamber with little topological framework of electricity and shearing force stimulation provided by the invention;
Fig. 2 is the sectional view that applies the flat flow chamber with little topological framework of electricity and shearing force stimulation provided by the invention;
Fig. 3 is the vertical view that applies the flat flow chamber with little topological framework of electricity and shearing force stimulation provided by the invention;
Fig. 4 is the upward view that applies the flat flow chamber with little topological framework of electricity and shearing force stimulation provided by the invention;
Fig. 5 is that the flat flow chamber with little topological framework of electricity and shearing force stimulation that applies shown in Figure 2 is along the sectional view of the A-A line among Fig. 2;
Fig. 6 is that the flat flow chamber with little topological framework of electricity and shearing force stimulation that applies shown in Figure 2 is along the sectional view of the B-B line among Fig. 2;
Fig. 7 is that the flat flow chamber with little topological framework of electricity and shearing force stimulation that applies shown in Figure 2 is along the sectional view of the C-C line among Fig. 2;
Fig. 8 is the vertical view that applies the flow cell bottom-side electrodes plate of electric and the flat flow chamber with little topological framework that shearing force stimulates provided by the invention;
Fig. 9 is that the flow cell bottom-side electrodes plate of electric and the flat flow chamber with little topological framework that shearing force stimulates that applies shown in Figure 8 is along the sectional view of the D-D line among Fig. 8;
Figure 10 is the vertical view of the flow cell bottom-side electrodes plate of the flat flow chamber with little topological framework that stimulates of the applied electricity that provides of the second embodiment of the present invention and shearing force;
Figure 11 is that the flow cell bottom-side electrodes plate of electric and the flat flow chamber with little topological framework that shearing force stimulates that applies shown in Figure 10 is along the sectional view of the E-E line among Figure 10;
Figure 12 is the sectional view of the flow cell bottom-side electrodes plate of the flat flow chamber with little topological framework that stimulates of the applied electricity that provides of the third embodiment of the present invention and shearing force;
Figure 13 is the side-view that applies the flow cell bottom-side electrodes plate of electric and the flat flow chamber with little topological framework that shearing force stimulates shown in Figure 12.
Wherein: the 1-conduit; The 2-upper flat plate; The 3-Upper gasket; The 4-diversion trench; The 5-middle plateform; The 6-rectangular slot; The 7-battery lead plate; The 8-wire; The 9-wire; The 10-lower gasket; Dull and stereotyped under the 11-; The 12-wire; The 13-wire; The 14-electrode; The 15-wire; The 16-battery lead plate; The 17-electrode; The 18-flow cell; The 19-cell; The 20-bolt; The 21-nut; The 22-thin slice; The 23-PDMS film; The 24-graphene film; The 25-wire.
Embodiment
Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.
Fig. 1 is the structural representation that applies the flat flow chamber with little topological framework of electricity and shearing force stimulation provided by the invention, its structure sectional view, top and bottom perspective views are respectively such as Fig. 2, Fig. 3 and shown in Figure 4, and Fig. 5, Fig. 6 and Fig. 7 are respectively the sectional views of A-A line among Fig. 2, B-B line and C-C line.
Embodiment 1:
By Fig. 1, Fig. 2, Fig. 3 and Fig. 4 as can be known, the flat flow chamber of implementing multi-direction electricity irritation of the present invention is comprised of three layer flat plate and the stack of two-layer pad, upper flat plate 2 is provided with fluid inlet and outlet, Upper gasket 3 is provided with diversion trench 4, middle plateform 5 arranges rectangular slot 6, and lower gasket 10 is provided with flow cell 18.Flow cell 18 width and length all should be greater than flow cell 18 thickness and thickness poor that is placed on battery lead plate 16 wherein.Upper flat plate 2, middle plateform 5 and lower dull and stereotyped 11 can be prepared by materials such as polycarbonate, resin glass or metals, and Upper gasket 3 and lower gasket 10 can be by preparations such as silicone resin.Such as Fig. 5, Fig. 6 and Fig. 7 as can be known, nutrient solution enters Upper gasket 3 intermediate cam shape diversion trenchs 4 by upper flat plate 2 upper conduits 1, so that fluid is fully dispersed and the effect that cushioned effectively reduces the flow-disturbing phenomenon of entrance.The nutrient solution of fully dispersing enters flow cell 18 by rectangular slot 6 on the middle plateform 5.
Described middle plateform 5 is provided with battery lead plate 7 above being positioned at flow cell 18.Battery lead plate 16 is placed on lower dull and stereotyped 11, is positioned at flow cell 18 belows.Pair of electrodes 14 and electrode 17 are placed respectively in flow cell 18 4 limits.Plant cells 19 on battery lead plate 16 surfaces carrying out flow cavity when experiment, perhaps experimentally need to show lining to battery lead plate 16 surfaces and plant again cell 19.Battery lead plate 7, electrode 14 and electrode 17 are made by graphene film or indium tin oxide conductive glass or indium tin oxide conductive film or the noble metal plate such as silver, gold or platinum.Middle plateform 5 electric pole plates 7 are by gluing connection, and the connection wire of battery lead plate 7 is drawn by the sulculus that is engraved on the middle plateform 5, and passes through sealant sealing.Be carved with sulculus on the lower gasket 10, be used for laying wire 8 and wire 25, wire 13 and wire 15, wire 9 and the wire 12 of connection electrode plate 16, electrode 14 and electrode 17, wire is used sealant sealing after putting into sulculus again.Upper flat plate 2, middle plateform 5, lower dull and stereotyped 11, Upper gasket 3 and lower gasket 10 same position places are processed with through hole, are used for wring bolt 20, by nut 21 compression fit.The number of bolt 20 can be symmetrical arranged according to the size of flat flow chamber 4-40.Hour can directly with the clamp pressing assembling, bolt 20 be set connect in the flat flow chamber size yet.
Fig. 8 is the vertical view that applies the flow cell bottom-side electrodes plate 16 of electric and the flat flow chamber with little topological framework that shearing force stimulates provided by the invention, and Fig. 9 is that flow cell bottom-side electrodes plate 16 shown in Figure 8 is along the sectional view of the D-D line among Fig. 8.Battery lead plate 16 comprises thin slice 22, PDMS film 23 and graphene film 24 among the figure, and the below will specifically introduce its making processes.
The silicon wafer that is 25~200mm at the thick 0.4~0.7mm of being diameter at first according to a conventional method forms micro-cannelure array, this little ditch groove width and deeply be respectively 100nm~20 μ m and 100nm~100 μ m, and spacing is 100nm~20 μ m between little groove.Prepare as follows PDMS film 23:
(1) mass ratio by 10: 1 of PDMS performed polymer and solidifying agent among the Sylgard184 is taken by weighing, be put in after fully mixing and vacuumize 20~30min in the vacuum drying oven, until bubble collapse.
(2) the PDMS mixed solution that mixes is poured into the silicon wafer surface that is placed in the plastic culture dish, form thick 100~1000 μ m PDMS films, be placed horizontally at and keep negative pressure-pumping 10min in the vacuum drying oven, colloid is fully contacted with silicon wafer template surface structure, and remove residual bubble.
(3) according to film thickness culture dish is placed horizontally at 80 ℃ of baking 1~2h in the air dry oven.Be cooled to room temperature after the curing, peel off the PDMS film that obtains surperficial little ditch, and be cut to desired shape with scalpel.
(4) the PDMS film is placed successively respectively ultrasonic cleaning 20min of 75% alcohol and deionized water, 90 ℃ of dry 1h of air dry oven.
(5) with the dilute hydrochloric acid immersion 2h of PDMS film with 1mol/L, improve surface hydrophilicity.Washed with de-ionized water, dry rear for subsequent use.
Then adopt the ultrasonic graphite method of peeling off of liquid phase to prepare the graphene dispersing solution that concentration is about 0.02~1.0mg/ml:
(1) Graphite Powder 99 is joined in DMF (N, N-Dimethylformamide, the DMF) solvent, process 1~2h (550~650W, 5s opens, the 5s pass) with the ultrasonic cell disrupte machine, leave standstill an evening.
(2) draw supernatant liquid and splash in the centrifuge tube, the centrifugal 30min of 5000rpm collects upper strata 4/5 liquid.
(3) gained liquid 7000rpm recentrifuge 30min collects the supernatant liquid body, obtains limpid transparent aterrimus graphene dispersing solution.
Then adopt spin-coating method (spin coating) to form graphene film 24 at PDMS film 23:
(1) prepares thin slice 22 with silicon chip or slide, PDMS film 23 is fixed on the thin slice 22, drip 0.1~1.0ml graphene dispersing solution in the center membrane position.
(2) leave standstill 5s, open the spin coating instrument, spin coating 10s under the rotating speed 1000rmp, then adjusting rotary speed is 3000rmp spin coating 30s.
(3) be positioned over drying in the vacuum drying oven, form the graphene film 24 of thick 1~50nm at PDMS film 23.
At the battery lead plate 16 two ends difference conducting wire 8 and the wire 25 that prepare, battery lead plate 16 and battery lead plate 7 can be used as pair of electrodes under study for action at last, also can be used as unitary electrode and use.Unite when using with battery lead plate 7, with in wire 8 or the wire 25 one or connect simultaneously power supply one utmost point; When using as unitary electrode, wire 8 is connected the two poles of the earth that connect respectively power supply with wire, allows electric current pass through 16 pairs of cells of battery lead plate 19 and produces electricity irritation.
Embodiment 2:
Little topological framework on the battery lead plate 16 of the present invention is not limited to little groove that embodiment 1 provides, and can also form at silicon wafer the micro-boss array of trilateral, tetragon, pentagon, hexagon or all kinds of shapes such as cylindrical.Take square boss as example, Figure 10 is the vertical view that can apply the flow cell bottom-side electrodes plate of electric and the flat flow chamber with little topological framework that shearing force stimulates, and Figure 11 is that the flow cell bottom-side electrodes plate of electric and the flat flow chamber with little topological framework that shearing force stimulates that applies shown in Figure 10 is along the sectional view of the E-E line among Figure 10.For obtaining the battery lead plate such as figure, can form square micro-boss array at silicon wafer first, this square micro-boss length of side is 100nm~20 μ m, and height is 100nm~100 μ m, and spacing is 100nm~20 μ m between micro-boss.Also can form the micro-boss array of other rules or irregular figure according to this method.And additive method is all identical with embodiment 1 with structure.
Embodiment 3:
Battery lead plate 16 of the present invention can be prepared into the flat board of consistency of thickness, realizes the hydrodynamic shear effect of homogeneous.Battery lead plate 16 also can be prepared into the clapboard that even thickness changes, and realizes continuous shear action.Figure 12 is the sectional view of the battery lead plate 16 of even thickness variation, and Figure 13 is the side-view of bottom-side electrodes plate 16 shown in Figure 12.When the battery lead plate 16 that the preparation even thickness changes, can keep PDMS film 23 thickness homogeneous, thin slice 22 is made the wedge shape of 5~60 ° of obliquitys, also can keep thin slice 22 thickness homogeneous, PDMS film 23 be made the wedge shape of 5~60 ° of obliquitys.For convenient processing, preferably keep PDMS film 23 thickness homogeneous, thin slice 22 is made the wedge shape of 5~60 ° of obliquitys.And additive method is all identical with embodiment 1 or embodiment 2 with structure.
The using method of flat flow chamber provided by the invention is as follows:
(1) select the battery lead plate 16 of required little topological framework according to the research needs, inoculating cell 19 on battery lead plate 16, plant cell 19 if inoculating cell 19 needs to show first lining to battery lead plate 16 surfaces again.
(2) after the battery lead plate 16 that will inoculate cell 19 is put into flow cell 18, by bolt or the good flat flow chamber of Fixture assembly.
(3) utilize teflon catheter that flow cavity, reservoir and peristaltic pump are connected into a system that circulates.
(4) select battery lead plate according to the research needs, the battery lead plate wire is connected with power supply.
(5) other devices beyond peristaltic pump, signal generator and the voltage stabilized source are all put into incubator.
(6) by peristaltic pump shearing force loading mode (continuously, vibration or pulse etc.) and intensity are set, by signal generator and voltage stabilized source electricity irritation binding mode (continuously, pulse etc.) and intensity are set.
(7) open peristaltic pump, signal generator and voltage stabilized source.After liquid in the reservoir is pumped by peristaltic pump, flow into flat flow chamber along conduit, the cell 19 on the battery lead plate 16 is applied shear action, then flow out, get back in the reservoir along flowing out conduit.If only need in three kinds of stimulation modes one or both, also can only open required equipment.When not needing to apply little topological framework stimulation, available common slide glass replaces battery lead plate 16.
(8) hormesis is closed peristaltic pump, signal generator and voltage stabilized source after finishing.
Flat flow chamber provided by the invention passes through at six face installing electrodes of flow cell plate, realization applies different mode and intensity electricity and hydrodynamic shear effect to cell, investigate simultaneously the little topological framework of substrate to the impact of characteristics of cell biology at the little topological framework of flow cell bottom-side electrodes plate surface formation, can also realize homogeneous hydrodynamic shear and continuous shear action by changing flow cell bottom-side electrodes plate thickness.Compared with prior art, flat flow chamber provided by the invention has following effect:
(1) flow cavity provided by the invention can to cell apply shearing force stimulation, electricity irritation and little topological framework in stimulating a kind of, two kinds and three kinds, be conducive to study and the response of observation of cell to the many factors combined stimulation, to realize acceleration of tissue repair speed and to improve the tissue repair quality.With respect to existing stimulation mode, can realize simultaneously in the device of a compact construction that many factors stimulates, and investigates the synergistic action effect of multifactor stimulation.
(2) flow cavity provided by the invention is convenient applies shearing force stimulation, electricity irritation and the stimulation of little topological framework, easier analog cell microenvironment in vivo to cell simultaneously.Existing apparatus can only realize that usually one or both factors stimulate, with cell in vivo microenvironment differ greatly.
(3) each stimulus all can realize various modes and intensity in the flow cavity provided by the invention.Wherein, shearing force loading mode and intensity can be by the peristaltic pump controls; Electrode stimulating pattern and intensity can be controlled by signal generator and voltage stabilized source, and can use simultaneously a plurality of electrodes to realize multi-direction electricity irritation; Little topological framework can be micro-cannelure array, also trilateral, tetragon, pentagon or hexagon micro-boss array, the also various topological structures such as cylindrical micro-boss array.Existing flow cavity device can not realize that substantially little topological framework stimulates, and the form of shearing force and electricity irritation is more single.
Just because of these composite factors, so that the flat flow chamber that flow cavity provided by the present invention is compared in the past has unique advantage.

Claims (9)

1. one kind can apply the flat flow chamber with little topological framework electric and that shearing force stimulates, formed by three layer flat plate and the stack of two-layer pad, upper flat plate is provided with fluid inlet and outlet, Upper gasket is provided with diversion trench, middle plateform arranges rectangular slot, lower gasket is provided with flow cell, flow cell six faces are equipped with battery lead plate, it is characterized in that: the described battery lead plate (16) that is positioned at flow cell (18) below comprises thin slice (22), PDMS film (23) and graphene film (24), PDMS film (23) is fixed on the thin slice (22) with different little topological frameworks, graphene film (24) forms by being spin-coated on PDMS film (23) surface, and battery lead plate (16) two ends are respectively arranged with wire (8) and wire (25).
2. the flat flow chamber with little topological framework that applies electricity and shearing force stimulation according to claim 1, it is characterized in that: the little topological framework on the described thin slice (22) is micro-cannelure array, this little groove and deeply be respectively 100nm~20 μ m and 100nm~100 μ m, spacing is 100nm~20 μ m between little groove.
3. the flat flow chamber with little topological framework that applies electricity and shearing force stimulation according to claim 1, it is characterized in that: the little topological framework on the described thin slice (22) is trilateral, tetragon, pentagon or hexagon micro-boss array, each limit length of side of this micro-boss is 100nm~20 μ m, height is 100nm~100 μ m, and spacing is 100nm~20 μ m between micro-boss.
4. the flat flow chamber with little topological framework that applies electricity and shearing force stimulation according to claim 1, it is characterized in that: the little topological framework on the described thin slice (22) is cylindrical micro-boss array, this micro-boss diameter is 100nm~20 μ m, height is 100nm~100 μ m, and spacing is 100nm~20 μ m between micro-boss.
5. the flat flow chamber with little topological framework that applies electricity and shearing force stimulation according to claim 1, it is characterized in that: described battery lead plate (16) is the clapboard of the flat board of consistency of thickness or even thickness variation, when the battery lead plate (16) that the preparation even thickness changes, can keep PDMS film (23) thickness homogeneous, thin slice (22) be made the wedge shape of 5~60 ° of obliquitys; Or keep thin slice (22) thickness homogeneous, PDMS film (23) is made the wedge shape of 5~60 ° of obliquitys.
6. the flat flow chamber with little topological framework that applies electricity and shearing force stimulation according to claim 1, it is characterized in that: described wire (8) is connected 25 with wire) one of or both connect simultaneously power supply one utmost point, battery lead plate (16) and battery lead plate (7) are used as pair of electrodes.
7. the flat flow chamber with little topological framework that applies electricity and shearing force stimulation according to claim 1, it is characterized in that: described wire (8) is connected 25 with wire) connect respectively two poles, battery lead plate (16) is used as an electrode separately, make electric current pass through battery lead plate (16) cell (19) is produced electricity irritation.
8. the flat flow chamber with little topological framework that applies electricity and shearing force stimulation according to claim 1 is characterized in that: described battery lead plate (7), electrode (14) and electrode (17) are made by graphene film or indium tin oxide conductive glass or indium tin oxide conductive film or silver, gold or platinum noble metals plate.
9. the flat flow chamber with little topological framework that applies electricity and shearing force stimulation according to claim 1, it is characterized in that: described thin slice (22) is made by silicon chip or slide.
CN201210417903.0A 2012-10-23 2012-10-23 Microtopological-structure plate flow chamber capable of applying electric and shearing force stimulation Active CN102936567B (en)

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