CN103805511B - The arteries simulation micro fluidic device directly can observed under high power objective - Google Patents
The arteries simulation micro fluidic device directly can observed under high power objective Download PDFInfo
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- CN103805511B CN103805511B CN201410054147.9A CN201410054147A CN103805511B CN 103805511 B CN103805511 B CN 103805511B CN 201410054147 A CN201410054147 A CN 201410054147A CN 103805511 B CN103805511 B CN 103805511B
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
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- C12M23/16—Microfluidic devices; Capillary tubes
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- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
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- 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
Abstract
The invention discloses a kind of arteries simulation micro fluidic device directly can observed under high power objective, this device is followed successively by transparent microchannel module (1), elastica (2), negative pressure generation module (3) and cover glass (10) from top to bottom; The bottom of described microchannel module 1 is provided with microchannel (8), flows for fluid; Described negative pressure generation module (3) inside is provided with runs through its upper and lower negative pressure hollow out groove (9), for generation of the negative pressure making elastica generation deformation; Thickness sum≤300 μm of described elastica (2) and cover glass (10).Micro fluidic device of the present invention not only can bear organ, the physiology of tissue, the pathological condition of hydrodynamic shear and mechanical stretching force in analogue body simultaneously, also directly can observe under high power objective, thus realize real-time monitored to the dynamic static change in individual cells fine structure and microchannel, provide effective tool more for related basic research and for the drug screening of arteriovascular diseases.
Description
Technical field
The present invention relates to biomedicine technical field, particularly relate to a kind of arteries simulation micro fluidic device directly can observed under high power objective.
Background technology
Abnormal blood flow kinetic factor is one of critical risk factor causing cardiovascular and cerebrovascular diseases, but its mechanism of action it be unclear that, and the limitation of traditional research method hinders Developments.In recent years, the foundation of the outer research model of vascular bodies and application facilitate the progress of correlative study greatly.
The haemodynamics in vitro study model of blood vessel produces mechanical stimulation kind when can flow through blood vessel according to they simulate blood is divided three classes, i.e. hydrodynamic shear model, Tensile stress model, hydrodynamic shear and Tensile stress interaction model.Hydrodynamic shear model mainly adopts laminar flow plate, and liquid applies fluid shear stress by the liquid-inlet and outlet being opened on both sides to the cell being planted in substrate; Tensile stress model then applies mechanical stretching stimulation by the deformation of elastica or plate to the superincumbent cell of adhesion.First two model can inquire into the Behavioral change of cell in single mechanical stimulation situation, but, be a complex environment having multiple mechanical stimulation in organism residing for cell, a cardiovascular systems in vitro study model closer to internal milieu must consider the effect of multiple mechanical stimulation to cell.
In recent years, along with the development of microflow control technique, people's designing and making micro fluidic device of various analogue body inner model:
The people such as Zhang (Zhang Q, Liu T, Qin J. (2012) A microfluidic-based device forstudy of transendothelial invasion of tumor aggregates in realtime.Lab Chip.Aug21; 12 (16): 2837-42) by external some important component being reconstructed biological blood vessel of microflow control technique, comprise lumen of vessels, endodermis, blood vessel surrounding substrate containing chemokine, be used for studying the adhesion of cancer cells aggregate and penetrate endodermis phagocytic process, but this device does not have the dynamics environment in analogue body well.
The people such as Srigunapalan (Srigunapalan S, Lam C, Wheeler AR, Simmons CA. (2011) A microfluidic membrane device to mimic critical components of the vascularmicroenvironment.Biomicrofluidics.5 (1): 13409.) made a device be made up of the PET film of a two PDMS passages and centre porous, this device can the key ingredient of external reconstruct blood vessel microenvironment, comprise hemodynamic shear-stress, the circulating cells factor, interaction between extracellular matrix protein and various kinds of cell, can be used for simulating nature and engineering blood vessel environment in study interaction between various kinds of cell matrix and cell-ECM.
The people such as Chen (Chen MB; Srigunapalan S; Wheeler AR, Simmons CA. (2013) A3Dmicrofluidic platform incorporating methacrylated gelatin hydrogels to studyphysiological cardiovascular cell-cell interactions.Lab Chip.Jul7; 13 (13): 2591-8.) device built may be used for the research such as drug screening of vasokinetic, valve biological study and vascular disease, the key feature of this device comprises fluid matasomatism on spatial disposition that various kinds of cell physiology is correlated with, endothelium monolayer, allow on the basis keeping cell to divide the interaction between different cell, gel-MA hydrogel be correlated with as physiology in subcutaneous 3D matrix maintain the long-term cultivation of cell.But these two devices can only provide hemodynamic shear-stress in research process, cannot the mechanical stretching force of blood vessel in analogue body.
The people such as Moore (Moore, J.E., Burki, E., Suciu, A., Zhao, S.M., Burnier, M., Brunner, H.R.and Meister, J.J. (1994) A Device for Subjecting VascularEndothelial-Cells to Both Fluid Shear-Stress and Circumferential Cyclic Stretch.AnnBiomed Eng.22,416-422) device that designs can apply hydrodynamic shear to cell simultaneously and drawing force stimulates.Method is by cell seeding at silicone tube internal surface, and at the import infusion fluid of silicone tube, liquid stream flows through cell surface, produces hydrodynamic shear to cell, and meanwhile, silicone tube can strut by the pressure of liquid miscarriage life, produces mechanical stretching force to cell.The feature of this device is can the dynamic process of beating of simulated blood vessel, but this device cannot carry out real-time monitored and intervention to the cell in this process.
The people such as Huh (Huh, D., Matthews, B.D., Mammoto, A., Montoya-Zavala, M., Hsin, H.Y.and Ingber, D.E. (2010) Reconstituting Organ-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, its purposes is simulation and research alveolar function, and its manufacture craft is relatively complicated, because film does not have upholder, easy distortion, not easily observation of cell form and change procedure under microscope.
The people such as Douville (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 mechanical stresses contribute to cell death anddetachment in a microfluidic alveolar model.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, liquid-gas interface can be formed, the microenvironment of simulated lung cystencyte.But above-mentioned micro fluidic device is only suitable for making alveolar model.
The people such as Zheng (Zheng W; Jiang B, Wang D, Zhang W; Wang Z, Jiang X. (2012) Amicrofluidic flow-stretch chip for investigating blood vessel biomechanics.Lab Chip.Sep21; 12 (18): 3441-50) device adopted may be used for applying separately hydrodynamic shear, separately applying mechanical stretching force to cell and applying the research of hydrodynamic shear and mechanical stretching force simultaneously, but can not observe kinetics of cells under high power lens.
Summary of the invention
For the defect of above-mentioned prior art, the object of the invention is to propose a kind ofly to provide hydrodynamic shear and balanced mechanical stretching force simultaneously, and the arteries simulation micro fluidic device of the Real-time and Dynamic observation to cell state and fine structure can be realized by high power objective.
For reaching this object, the present invention by the following technical solutions:
First aspect, the invention provides a kind of arteries simulation micro fluidic device directly can observed under high power objective, this device comprises four transparent parts: microchannel module (1), the elastica (2) suitable with it, negative pressure generation module (3), and cover glass (10);
Described four parts press from top to bottom microchannel module (1), elastica (2), negative pressure generation module (3), cover glass (10) the superimposed successively and covalent bonding of order together;
The bottom of described microchannel module (1) is provided with microchannel (8), flows for fluid; Described microchannel (8) is with elastica (2) for bottom, and the top of described microchannel (8) is provided with the fluid intake (4) and fluid outlet (5) that communicate with outside;
Described negative pressure generation module (3) inside is provided with runs through its upper and lower negative pressure hollow out groove (9), for generation of the negative pressure making elastica (2) that deformation occur; Described negative pressure hollow out groove (9) is positioned at the below of microchannel (8), and horizontal sectional area is greater than microchannel (8), at the top of described negative pressure hollow out groove (9) and be positioned at microchannel (8) outside both sides be respectively equipped with the first gas circuit opening (6) and the second gas circuit opening (7) that communicate with outside;
Thickness sum≤300 μm of described elastica (2) and cover glass (10).
In above-mentioned arteries simulation micro fluidic device, as preferably, described covalent bonding is realized by plasma oxidation process.
As preferably, described fluid intake (4) and fluid outlet (5) run through respectively by the PE pipe suitable with it, preferred vertical runs through microchannel module (1), communicate, be preferably connected with cell culture medium drive system with outside;
Preferably, described fluid intake (4) and fluid outlet (5) are circular port, and more preferably, the diameter of described circular port is 0.5mm.
As preferably, described first gas circuit opening (6) and the second gas circuit opening (7) run through respectively by the PE pipe suitable with it, preferred vertical runs through negative pressure hollow out groove (9) top elastica (2) and microchannel module (1), communicate with outside, be preferably connected with negative pressure generator;
Preferably, described first gas circuit opening (6) and the second gas circuit opening (7) are circular port, and more preferably, the diameter of described circular port is 0.5mm.
As preferably, described microchannel module (1), elastica (2) and negative pressure generation module (3) are made by polydimethylsiloxane (PDMS) material; Preferably, described elastica (2) surface is smooth or has nanotopology.
As preferably; the top elastica of described negative pressure hollow out groove (9) and bottom cover slide not bonding, be filled with liquid lubricant therebetween but be not full of, and makes it can move each other under suction function; and when there is relative movement, elastica (2) is played to the effect of lubrication and protection.
As preferably, the length of described microchannel module (1), elastica (2), negative pressure generation module (3) is 28-35mm, and width is 20-25mm; The thickness of described microchannel module (1) is 2-6mm, preferably 3-5mm; The thickness of described elastica (2) is 10-100 μm, preferred 50-100 μm; The thickness of described negative pressure generation module (3) is 0.3-0.5mm; The thickness of described cover glass (10) is 150-200 μm; Thickness sum≤280 μm of described elastica (2) and cover glass (10);
Further preferably, the length of described microchannel (8) is 20-23mm, and width is 2-3mm, is highly 200-300 μm; The length of described negative pressure hollow out groove (9) is 25-28mm, and width is 2.5-3.5mm, is highly 0.3-0.5mm.
In a specific embodiments of the present invention, the length of described microchannel module (1), elastica (2), negative pressure generation module (3) is 28mm, and width is 21.5mm; The thickness of described microchannel module (1) is 4mm; The thickness of described elastica (2) is 100 μm; The thickness of described negative pressure generation module (3) is 0.4mm; The thickness of described cover glass (10) is 170 μm; The length of described microchannel (8) is 20mm, and width is 2mm, is highly 200 μm; The length of described negative pressure hollow out groove (9) is 25mm, and width is 3mm, is highly 0.3mm.
Above-mentioned arteries simulation micro fluidic device can apply hydrodynamic shear and mechanical stretching force simultaneously, and Real-time and Dynamic observes cell state and fine structure thereof under high power objective, and its functional realiey process is as follows:
First by the corresponding pipeline of the fluid intake (4) of microchannel (8), cell suspension is added microchannel (8), 37 degrees Celsius, under 5% carbon dioxide conditions, make cell attachment in the elastica (2) of channel bottom, then, the input terminus of microchannel (8) (i.e. fluid intake (4) and the PE pipe that connects thereof) and output terminal (namely fluid outlet (5) and the PE that is connected thereof manage) are connected with cell culture medium drive system.First gas circuit opening (6) of negative pressure generation module (3) is connected with negative pressure generator by PE pipe with the second gas circuit opening (7).Then, start substratum drive system and negative pressure generator simultaneously, substratum drive system can drive the liquid-flow in microchannel, thus hydrodynamic shear is produced to the cell that substrate in passage is adhered to, and the negative pressure that negative pressure generator produces can be conducted in the negative pressure hollow out groove (9) in negative pressure generation module (3) by PE pipe, make the elastica (2) of negative pressure hollow out groove (9) top that symmetrical deformation occur, thus pull the elastica bottom microchannel, make it to occur: the i) deformation of horizontal direction, thus make the superincumbent cell of attaching be subject to mechanical stretching force, ii) deformation straight down, makes its cover glass being close to negative pressure hollow out channel bottom (10) surface, is convenient to high power objective and observes.
Because elastica (2) is to be parallel to the deformation on passage (8) long axis direction very little, negligible, be applied to elastica (2) and elastica (2) if upper cell advocate perpendicular to passage (8) long axis direction, therefore, the stressing conditions of cell is: be parallel to the hydrodynamic shear in passage (8) direction and the mechanical stretching force perpendicular to passage (8) direction.Due to elastica (2) and total thickness≤300 μm of cover glass (10), so when microchannel (8) bottom elastica (2) pull down extend close to or arrive cover glass (10) time, high power objective can be used directly to observe cell state in elastica (2) and fine structure thereof, thus realize the Real-time and Dynamic observation of cell state and fine structure thereof.
Second aspect, the invention provides the purposes of arteries simulation micro fluidic device as described in relation to the first aspect, it is characterized in that, for pathomechanism research and/or the drug screening of arteriovascular diseases; Preferably, for the preparation of the pathomechanism research of arteriovascular diseases or the biological detection reagent kit of drug screening.
The third aspect, the invention provides the biological detection reagent kit that a kind of arteries comprised described in first aspect simulates micro fluidic device; Preferably, described biological detection reagent kit also comprises detection reagent and damping fluid; Further preferably, described detection reagent is vasoactive small molecules, cytokine, antibody or the medicine for screening.
Arteries simulation micro fluidic device of the present invention has following Characteristics and advantages:
1, microchannel, elastica, negative pressure generation module three part-structure and high integration functionally, and all parts all make of optically transparent material, are easy to the situation in naked eyes or Microscopic observation passage.
2, volume is little, and structure is simple, is easy to make and use.
3, the long-term cultivation of cell can be realized in incubator.
4, in cell cultivation process, the chemical micro-environment of cultivation can be changed on demand.
5, can under high power objective the fine structure of direct observing device inner cell, and can realize observing the dynamic realtime of cell in experimentation.
6, can in a device separately or comprehensively apply mechanical stretching force and hydrodynamic shear, and can the size of flexible modulation hydrodynamic shear and the frequency of mechanical stretching force, with the mechanical environment under different positions in simulated blood vessel or pathological conditions.
There is the cell of many Organ and tissues to be subject to the stimulation of hydrodynamic shear and mechanical stretching force in body simultaneously, the physiological and pathological research of these organs lacks the external model of suitable convenient operation and real-time monitored directly perceived at present, the invention of this device is that relevant research provides platform, effectively can promote the progress of correlative study.
Arteries of the present invention simulation micro fluidic device not only can bear the organ of hydrodynamic shear and mechanical stretching force in analogue body simultaneously, the physiology of tissue, pathological condition, set up the external model of disease in body, for the pathological study of disease, drug screening and nano particle is modified carries the research work such as gene transfecting cell and provide manipulation in vitro platform, also directly can observe under high power objective, thus the real-time monitored realized the dynamic static change in individual cells fine structure and microchannel, effective tool is more provided for related basic research and for the drug screening of arteriovascular diseases.
To sum up, the present invention is applicable to all physiology or Pathological experiment of needing cell is applied separately to the physiology of hydrodynamic shear or mechanical stretching force or Pathological experiment or applies hydrodynamic shear or mechanical stretching force simultaneously, particularly can be the platform that the long-time Real Time Observation of cell under mechanical stimulation and dynamic intervention experiment provide convenient operation and real-time monitored, and provide experiment porch for the fine structure of long-term real-time monitored cell under mechanical stimulation; In addition, also while hydrodynamic shear and/or mechanical stretching force are applied to cell, chemical stimulation can be applied, thus provide platform for the medicine screening of arteriovascular diseases.
Accompanying drawing explanation
Fig. 1 is the structural representation of arteries of the present invention simulation micro fluidic device; Wherein, 1 is microchannel module, and 2 is elastica, and 3 is negative pressure generation module, and 4 is fluid intake, and 5 is fluid outlet, and 6 is the first gas circuit opening, and 7 is the second gas circuit opening, and 8 is microchannel, and 9 is negative pressure hollow out groove, and 10 is cover glass;
Fig. 2 is that in arteries of the present invention simulation micro fluidic device, elastica carries out the experimental result schematic diagram stretched under negative pressure; Wherein A is the elastica before stretching, and B is the elastica after stretching;
Fig. 3 is the cell fine structure in the arteries simulation micro fluidic device of the present invention of 63 times of thing Microscopic observations.
Embodiment
Technical scheme of the present invention is further illustrated by embodiment below in conjunction with accompanying drawing.
Fig. 1 is the structural representation of arteries of the present invention simulation micro fluidic device.As shown in Figure 1, this device comprises transparent microchannel module (1) and the transparent elastica (2) suitable with it and negative pressure generation module (3), and transparent cover glass (10).Press from top to bottom microchannel module (1), elastica (2), negative pressure generation module (3), cover glass (10) order superimposed successively, and through plasma oxidation process covalent bonding together, described microchannel module (1), elastica (2), negative pressure generation module (3) are made by polydimethylsiloxane (PDMS) material.The length of described microchannel module (1), elastica (2), negative pressure generation module (3) is 28mm, and width is 21.5mm; The thickness of described microchannel module (1) is 4mm; The thickness of described elastica (2) is 100 μm; The thickness of described negative pressure generation module (3) is 0.4mm; The thickness of described cover glass (10) is 170 μm.The length of described microchannel (8) is 20mm, and width is 2mm, is highly 200 μm.The length of described negative pressure hollow out groove (9) is 25mm, and width is 3mm, is highly 0.3mm.Not bonding between the top elastica (2) of described negative pressure hollow out groove (9) and bottom cover slide (10); a small amount of liquid lubricant (but not being full of) is filled in negative pressure hollow out groove (9); make it can move each other under suction function; and when relative movement occurs elastica (2), elastica (2) is played to the effect of lubrication and protection.Described fluid outlet (5) and fluid intake (4) are the circular port that diameter is 0.5mm, be symmetrically set in the both sides at microchannel (8) top, described fluid outlet (5) and fluid intake (4) vertically run through described microchannel module by managing with its suitable PE, are connected with cell culture medium drive system.Described first gas circuit opening (6) and the second gas circuit opening (7) are also the circular port that diameter is 0.5mm, be symmetrically set in described negative pressure hollow out groove (9) top and be positioned at microchannel (8) outside both sides, described first gas circuit opening (6) and the second gas circuit opening (7) run through elastica (2) and the microchannel module (1) of negative pressure hollow out groove (9) top by the PE pipe suitable with it, are connected with negative pressure generator.
During use, first by the fluid intake (4) of microchannel (8) corresponding pipeline, cell suspension is added microchannel (8), 37 degrees Celsius, under 5% carbon dioxide conditions, make cell attachment in the elastica of channel bottom, then, the fluid intake (4) of microchannel is connected with cell culture medium drive system by its corresponding PE pipe with fluid outlet (5), then the first gas circuit opening (6) of negative pressure generation module is connected with negative pressure generator with the second gas circuit opening (7).Then, start substratum drive system and negative pressure generator simultaneously, substratum drive system can drive the liquid-flow in microchannel, thus hydrodynamic shear is produced to the cell that passage (8) interior substrate is adhered to, and the negative pressure that negative pressure generator produces can be conducted in the negative pressure hollow out groove (9) in negative pressure generation module (3) by PE pipe, make the elastica (2) of negative pressure hollow out groove (9) top that symmetrical deformation occur, thus pull the elastica bottom microchannel, make it to occur: the i) deformation of horizontal direction, thus make the superincumbent cell of attaching be subject to mechanical stretching force, ii) deformation straight down, makes its cover glass being close to negative pressure hollow out channel bottom (10) surface, is convenient to high power objective and observes.
Because elastica (2) is to be parallel to the deformation on passage (8) long axis direction very little, negligible, be applied to elastica (2) and elastica (2) if upper cell advocate perpendicular to passage (8) long axis direction, therefore, the stressing conditions of cell is: be parallel to the hydrodynamic shear in passage (8) direction and the mechanical stretching force perpendicular to passage (8) direction.Due to elastica (2) and total thickness≤300 μm of cover glass (10), so when microchannel (8) bottom elastica (2) pull down extend close to or arrive cover glass (10) time, high power objective can be used directly to observe cell state in elastica (2) and fine structure thereof, thus realize the Real-time and Dynamic observation of cell state and fine structure thereof.
When embodiment 1 arteries simulation of the present invention micro fluidic device runs, the stretching of elastica detects
HeLa cell is added the microchannel in arteries of the present invention simulation micro fluidic device, 37 DEG C, under 5% carbon dioxide conditions, make cell attachment in elastica; Then stretch under suction function, the cell in 20 times of thing Microscopic observation elasticas, as shown in Figure 2, the elastica after negative pressure stretches is compared with before stretching, and extensibility is about 8% for result.
The cell fine structure example in arteries of the present invention simulation micro fluidic device is observed under embodiment 2 high power objective
Human umbilical vein endothelial cells is added the microchannel in arteries of the present invention simulation micro fluidic device, 37 DEG C, under 5% carbon dioxide conditions, make cell attachment in elastica; Simulate arterial vascular mechanical condition and carry out cell cultures after 2 hours, cell is fixed, and nucleus and microfilament are dyeed, under the condition that elastica stretches, the fine structure of cell in 63 times of sem observation elasticas, result as shown in Figure 3, clearly can observe the fine structure of cell.
Applicant states, the present invention illustrates micro fluidic device of the present invention by above-mentioned drawings and Examples, but the present invention is not limited to above-mentioned drawings and Examples.Person of ordinary skill in the field should understand, any improvement in the present invention, all drops within protection scope of the present invention and open scope.
Claims (20)
1. the arteries simulation micro fluidic device directly can observed under high power objective, this device comprises four transparent parts: microchannel module (1), the elastica (2) suitable with it, negative pressure generation module (3), and cover glass (10);
Described four parts press from top to bottom microchannel module (1), elastica (2), negative pressure generation module (3), cover glass (10) the superimposed successively and covalent bonding of order together;
The bottom of described microchannel module (1) is provided with microchannel (8), flows for fluid; Described microchannel (8) is with elastica (2) for bottom, and the top of described microchannel (8) is provided with the fluid intake (4) and fluid outlet (5) that communicate with outside;
Described negative pressure generation module (3) inside is provided with runs through its upper and lower negative pressure hollow out groove (9), for generation of the negative pressure making elastica (2) that deformation occur; Described negative pressure hollow out groove (9) is positioned at the below of microchannel (8), and horizontal sectional area is greater than microchannel (8), at the top of described negative pressure hollow out groove (9) and be positioned at microchannel (8) outside both sides be respectively equipped with the first gas circuit opening (6) and the second gas circuit opening (7) that communicate with outside;
Thickness sum≤300 μm of described elastica (2) and cover glass (10);
Described fluid intake (4) and fluid outlet (5) are circular port, and the diameter of described circular port is 0.5mm;
Described first gas circuit opening (6) and the second gas circuit opening (7) are circular port, and the diameter of described circular port is 0.5mm.
2. arteries simulation micro fluidic device according to claim 1, it is characterized in that, described covalent bonding is realized by plasma oxidation process.
3. arteries simulation micro fluidic device according to claim 1, it is characterized in that, described fluid intake (4) and fluid outlet (5) run through microchannel module (1) respectively by the PE pipe suitable with it, communicate with outside.
4. arteries simulation micro fluidic device according to claim 3, it is characterized in that, described fluid intake (4) and fluid outlet (5) vertically run through microchannel module (1) respectively by managing with its suitable PE, communicate with outside.
5. arteries simulation micro fluidic device according to claim 3, it is characterized in that, described fluid intake (4) and fluid outlet (5) vertically run through microchannel module (1) respectively by managing with its suitable PE, are connected with cell culture medium drive system.
6. arteries simulation micro fluidic device according to claim 3, it is characterized in that, described first gas circuit opening (6) and the second gas circuit opening (7) run through elastica (2) and the microchannel module (1) of negative pressure hollow out groove (9) top respectively by the PE pipe suitable with it, communicate with outside.
7. arteries simulation micro fluidic device according to claim 6, it is characterized in that, described first gas circuit opening (6) manages the vertical elastica (2) and the microchannel module (1) that run through negative pressure hollow out groove (9) top with the second gas circuit opening (7) respectively by with its suitable PE, communicates with outside.
8. arteries simulation micro fluidic device according to claim 6, it is characterized in that, described first gas circuit opening (6) and the second gas circuit opening (7) run through elastica (2) and the microchannel module (1) of negative pressure hollow out groove (9) top respectively by the PE pipe suitable with it, are connected with negative pressure generator.
9. arteries simulation micro fluidic device according to claim 1, it is characterized in that, described microchannel module (1), elastica (2) and negative pressure generation module (3) are made by polydimethyl siloxane material.
10. arteries simulation micro fluidic device according to claim 1, it is characterized in that, described elastica (2) surface is smooth or has nanotopology.
11. arteries simulation micro fluidic devices according to claim 1, it is characterized in that, the top elastica of described negative pressure hollow out groove (9) and bottom cover slide not bonding, be filled with liquid lubricant therebetween.
12. arteries simulation micro fluidic devices according to claim 1, it is characterized in that, the length of described microchannel module (1), elastica (2), negative pressure generation module (3) is 28-35mm, and width is 20-25mm; The thickness of described microchannel module (1) is 2-6mm; The thickness of described elastica (2) is 10-100 μm; The thickness of described negative pressure generation module (3) is 0.3-0.5mm; The thickness of described cover glass (10) is 150-200 μm; Thickness sum≤280 μm of described elastica (2) and cover glass (10).
13. arteries simulation micro fluidic devices according to claim 12, it is characterized in that, the thickness of described microchannel module (1) is 3-5mm; The thickness of described elastica (2) is 50-100 μm.
14. arteries simulation micro fluidic devices according to claim 1, it is characterized in that, the length of described microchannel (8) is 20-23mm, and width is 2-3mm, is highly 200-300 μm; The length of described negative pressure hollow out groove (9) is 25-28mm, and width is 2.5-3.5mm, is highly 0.3-0.5mm.
15. arteries simulation micro fluidic devices according to claim 12, it is characterized in that, the length of described microchannel module (1), elastica (2), negative pressure generation module (3) is 28mm, and width is 21.5mm; The thickness of described microchannel module (1) is 4mm; The thickness of described elastica (2) is 100 μm; The thickness of described negative pressure generation module (3) is 0.4mm; The thickness of described cover glass (10) is 170 μm; The length of described microchannel (8) is 20mm, and width is 2mm, is highly 200 μm; The length of described negative pressure hollow out groove (9) is 25mm, and width is 3mm, is highly 0.3mm.
16. as described in any one of claim 1-15 arteries simulation micro fluidic device purposes, it is characterized in that, for arteriovascular diseases pathomechanism research and/or drug screening.
17. purposes according to claim 16, is characterized in that, for the preparation of the pathomechanism research of arteriovascular diseases or the biological detection reagent kit of drug screening.
18. 1 kinds of arteries comprised according to any one of claim 1-15 simulate the biological detection reagent kit of micro fluidic device.
19. biological detection reagent kits according to claim 18, is characterized in that, described biological detection reagent kit also comprises detection reagent and damping fluid.
20. biological detection reagent kits according to claim 19, is characterized in that, described detection reagent is vasoactive small molecules, cytokine, antibody or the medicine for screening.
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| CN109239319A (en) * | 2018-10-31 | 2019-01-18 | 清华大学 | It is a kind of for the circulation device containing micron order hollow channel gel rubber material |
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| CN111208282B (en) * | 2020-01-14 | 2021-12-24 | 安徽奇书生物科技有限公司 | Multi-channel mixed flow injection type perfusion microscope |
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