CN117229917A - Pump-driving-free organoid chip - Google Patents
Pump-driving-free organoid chip Download PDFInfo
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- CN117229917A CN117229917A CN202311524199.3A CN202311524199A CN117229917A CN 117229917 A CN117229917 A CN 117229917A CN 202311524199 A CN202311524199 A CN 202311524199A CN 117229917 A CN117229917 A CN 117229917A
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- 210000002220 organoid Anatomy 0.000 title claims abstract description 102
- 239000007788 liquid Substances 0.000 claims abstract description 107
- 239000001963 growth medium Substances 0.000 claims abstract description 65
- 238000001704 evaporation Methods 0.000 claims abstract description 54
- 230000008020 evaporation Effects 0.000 claims abstract description 48
- 239000012528 membrane Substances 0.000 claims abstract description 7
- 239000002609 medium Substances 0.000 claims description 18
- 239000012530 fluid Substances 0.000 claims description 10
- 239000012780 transparent material Substances 0.000 claims description 6
- 239000004793 Polystyrene Substances 0.000 claims description 3
- 239000004205 dimethyl polysiloxane Substances 0.000 claims description 3
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims description 3
- 229920003229 poly(methyl methacrylate) Polymers 0.000 claims description 3
- -1 polydimethylsiloxane Polymers 0.000 claims description 3
- 239000004926 polymethyl methacrylate Substances 0.000 claims description 3
- 239000004033 plastic Substances 0.000 claims description 2
- 229920003023 plastic Polymers 0.000 claims description 2
- 229920002223 polystyrene Polymers 0.000 claims description 2
- 238000009834 vaporization Methods 0.000 claims 1
- 230000008016 vaporization Effects 0.000 claims 1
- 230000010412 perfusion Effects 0.000 abstract description 9
- 230000000694 effects Effects 0.000 abstract description 3
- 238000000034 method Methods 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 10
- 239000003814 drug Substances 0.000 description 6
- 230000036770 blood supply Effects 0.000 description 5
- 230000009471 action Effects 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 3
- 238000012377 drug delivery Methods 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 238000001746 injection moulding Methods 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 230000017531 blood circulation Effects 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000010874 in vitro model Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
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- 238000001338 self-assembly Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
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- 210000001519 tissue Anatomy 0.000 description 2
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- 206010028980 Neoplasm Diseases 0.000 description 1
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- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000003255 drug test Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
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- 210000000056 organ Anatomy 0.000 description 1
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention relates to the field of biotechnology, and discloses a pump-driven organoid chip which comprises a top layer, a storage layer, a middle layer, a culture medium layer, a culture room layer and a bottom layer which are sequentially overlapped from top to bottom; the storage layer is provided with a culture medium storage chamber and a liquid evaporation chamber communicated with the outside; the surface of the outlet of the liquid evaporation chamber is covered with a microporous filter membrane; the culture chamber layer is provided with a plurality of organoid culture chambers, and the organoid culture chambers are communicated with a liquid inlet of the organoid culture chamber arranged on the top layer; the liquid inlet of the organoid culture chamber sequentially passes through the storage layer, the middle layer and the culture medium layer to be communicated with the organoid culture chamber; the culture medium layer is provided with a culture medium channel which is communicated with the liquid evaporation chamber, the culture medium storage chamber and the organoid culture chamber; the invention gets rid of the dependence on any liquid driving force equipment, can provide driving force for the flow of the culture medium in the chip by virtue of the liquid self-evaporation effect and the principle of the communicating vessel, and can realize dynamic 3D perfusion culture by the single organoid chip.
Description
Technical Field
The invention relates to the technical field of organoid chips, in particular to a pump-driving-free organoid chip.
Background
With the development of cell biology and organoid technology, cell three-dimensional culture technology is gradually replacing the traditional two-dimensional cell culture technology. At present, various types of cells have stronger self-assembly capability, such as pluripotent stem cells, tumor cells, tissue cells and the like. Three-dimensional cytoballs are three-dimensional aggregates formed by self-assembly of various cells, more approximate to the structural morphology of tissue cells in vivo and more beneficial to the research of the functional mechanism thereof. Three-dimensional cytospheres can therefore be used in a wide variety of biological and biomedical fields of research, such as development, pathology, pharmacology, cancer therapy, and the like.
In recent years, organ-chip technology provides an in vitro model for studying the functions and mechanisms of different organs, and is also widely applied to drug or therapeutic regimen screening. The organoid chip combines the front technologies of the two life sciences and engineering fields, and the constructed high-throughput and high-bionic in-vitro model construction platform has wide application in the fields of new medicine research and development, disease modeling, personalized accurate medical treatment and the like. At present, the organoid culture and the drug experiments are basically carried out in a culture dish or an orifice plate, and cannot simulate the microenvironment for the survival of cells in a human body. Second, it is not possible to mimic continuous natural flow drug delivery conditions in an in vivo blood supply environment while observing the ability of organoid-related responses.
The prior art discloses a microfluidic system for a drug test for tumor organoid culture and a use method thereof, wherein the microfluidic system comprises two pumps for providing flow power, two microfluidic chips are connected in series, and a first microfluidic chip is used for establishing different drug concentration gradient environments. The second chip is used for continuously pouring the medicine-containing culture medium into the organoids, and finally a waste liquid collector is connected to collect waste liquid. Preferably simulates the continuous natural flow drug delivery process in the blood supply environment in the body and observes the relevant response capability of the organoids to the drugs in real time. However, the whole system is complex, the system construction process is complex, the pump is required to provide flow power, and the occupied space is large. An organoid culture chip and method of use is disclosed as application No. 2020108923373. The chip is a nested structure which can be placed in a common cell culture dish and a pore plate, a plurality of micropores are arranged at the bottom of the chip, after cells are inoculated to the chip, under the combined action of gravity and mechanical force generated by the inclined side wall, the cells are gathered at the bottom of the micropores, three-dimensional cytoballs are spontaneously formed, and the formed cytoballs can be further cultured to form organoid tissues. The chip in the method can only perform static culture of the organoid, does not support dynamic perfusion culture, can not simulate continuous and natural flowing drug delivery conditions in a blood supply environment in vivo, and can not observe the relevant response capability of the organoid to continuous and dynamic perfusion administration.
Disclosure of Invention
The invention provides a pump-free organoid chip which aims at the problems existing in the prior art.
The technical scheme adopted by the invention is as follows: a pump-free organoid chip comprises a top layer, a storage layer, a middle layer, a culture medium layer, a culture room layer and a bottom layer which are sequentially stacked into an integral structure from top to bottom;
the storage layer is provided with a culture medium storage chamber and a liquid evaporation chamber communicated with the outside; the surface of the outlet of the liquid evaporation chamber is covered with a microporous filter membrane with the diameter of 0.22 mu m;
the culture chamber layer is provided with a plurality of organoid culture chambers, and the organoid culture chambers are communicated with a liquid inlet of the organoid culture chamber arranged on the top layer; the liquid inlet of the organoid culture chamber sequentially passes through the storage layer, the middle layer and the culture medium layer to be communicated with the organoid culture chamber;
the culture medium layer is provided with a culture medium channel which is communicated with the liquid evaporation chamber, the culture medium storage chamber and the organoid culture chamber.
Further, the organoid culture chambers are arranged in an array.
Further, the organoid culture chamber is communicated with a liquid inlet of the organoid culture chamber through a liquid inlet channel of the organoid culture chamber; the liquid inlet channel of the organoid culture chamber is of a Y-shaped bifurcation structure.
Furthermore, the organoid culture chamber is provided with two rows, and the liquid inlet of the organoid culture chamber is provided with two liquid inlets for the organoid culture chamber of the two rows respectively.
Further, the culture medium channel is communicated with the culture medium storage chamber through a liquid outlet of the culture medium storage chamber arranged in the middle layer; the culture medium channel is communicated with the liquid evaporation chamber through a liquid inlet of the liquid evaporation chamber arranged in the middle layer.
Further, the culture medium channel main body is of a rectangular structure, and two ends of the culture medium channel main body are of triangular structures to form a hexagonal structure; one end is communicated with a liquid outlet of the culture medium storage chamber, and the other end is communicated with a liquid inlet of the liquid evaporation chamber.
Further, a liquid inlet of the culture medium storage chamber and a liquid evaporation chamber outlet are formed in the top layer.
Further, the area of the outlet of the liquid evaporation chamber is 0.32 cm 2 ～4.5 cm 2 。
Further, the top layer, the storage layer, the middle layer, the culture medium layer, the culture chamber layer and the bottom layer are made of transparent materials, and one of polydimethylsiloxane, polystyrene plastic and polymethyl methacrylate is adopted.
The beneficial effects of the invention are as follows:
(1) According to the invention, through the evaporation effect on the culture medium in the liquid evaporation chamber, the liquid amount in the liquid evaporation chamber is reduced, and the driving force can be provided for the movement of fluid in the chip under the action of the communicating vessel principle, so that the liquid in the culture medium storage chamber slowly and continuously flows to the liquid evaporation chamber, and the nutrient substances of the organoid culture in the organoid culture chamber are updated and supplemented by the culture medium flowing through the culture medium channel;
(2) The invention gets rid of the dependence on any liquid driving force equipment, and can provide driving force for the flow of the culture medium in the chip by virtue of the principle of liquid self-evaporation and a communicating vessel; the continuous and natural flowing blood circulation process in the blood supply environment in the body is simulated, and the update and the supplement of the culture medium for the organoid in the organoid culture chamber are realized;
(3) According to the invention, the control of liquid flow can be realized through the evaporation surface areas with different sizes, the dependence of the chip on equipment is eliminated, and the dynamic 3D perfusion culture can be realized by the single organoid chip.
Drawings
FIG. 1 is a schematic view of the structure of the device of the present invention.
In the figure: 1-top layer, 101-medium reservoir liquid inlet, 102-organoid culture chamber liquid inlet, 103-liquid evaporation chamber outlet, 2-reservoir, 201-medium reservoir, 202-liquid evaporation chamber, 3-middle layer, 301-medium reservoir liquid outlet, 302-liquid evaporation chamber liquid inlet, 4-medium layer, 401-medium channel, 5-culture chamber layer, 501-organoid culture chamber, 502-organoid culture chamber liquid inlet channel, 6-bottom layer, 7-microporous filter membrane.
Detailed Description
The invention will be further described with reference to the drawings and specific examples.
As shown in FIG. 1, the pump-free organoid chip comprises a top layer 1, a storage layer 2, a middle layer 3, a culture medium layer 4, a culture chamber layer 5 and a bottom layer 6 which are sequentially stacked into a whole structure from top to bottom; the layers in the invention are arranged for the convenience of expression and manufacturing, and in practical application, the layers are combined into a whole structure, and the whole structure can be directly prepared by injection molding and other methods.
The reservoir 2 is provided with a medium reservoir 201 and a liquid evaporation chamber 202 communicating with the outside; the surface of the outlet 103 of the liquid evaporation chamber is covered with a microporous filter membrane 7 with the thickness of 0.22 mu m; the size of the microporous filter membrane 7 is selected for filtering and sterilizing, so that the pollution to the organoid culture caused by the chip carried out by the external microorganisms is isolated, and the area of the microporous filter membrane 7 can completely cover the outlet 103 of the liquid evaporation chamber.
The culture chamber layer 5 is provided with a plurality of organoid culture chambers 501, and the organoid culture chambers 501 are communicated with the organoid culture chamber liquid inlet 102 arranged on the top layer 1; the liquid inlet 102 of the organoid culture chamber sequentially passes through the storage layer 2, the middle layer 3 and the culture medium layer 4 to be communicated with the organoid culture chamber 501;
the culture substrate layer 4 is provided with a culture medium channel 401, and the culture medium channel 401 communicates with the liquid evaporation chamber 202, the culture medium storage chamber 201 and the organoid culture chamber 501.
The culture medium storage chamber 201 can be provided with bottom surfaces and thicknesses of different sizes according to the culture time requirements, different volume capacities are realized, the bottom surface shape of the culture medium storage chamber is preferably round-corner rectangle or round, the round-corner structure is beneficial to the liquid to fully fill the space of the culture medium storage chamber 201 along with the tension, and dead space is not easy to generate; two liquid inlets 101 of the corresponding culture medium storage chamber are arranged, and if the liquid inlets are of a round-corner rectangular structure, the two liquid inlets are arranged at corresponding positions of two diagonal end points of the culture medium storage chamber 201; if the liquid inlet is circular, the two liquid inlets are respectively arranged at the positions corresponding to the diameter vertexes. The bottom surface of the liquid evaporation chamber 202 is preferably circular, and the diameter of the circle can be adjusted according to the liquid perfusion flow rate required by culture, so that the evaporation surface area can be adjusted to adjust the liquid evaporation rate and evaporation amount, and further adjust the liquid flow rate in the culture medium channel.
The organoid culture chambers 501 are arranged in an array, and the organoid culture chambers 501 are preferably circular, so that the injected liquid is beneficial to filling the whole culture chamber, and dead space is avoided; the number of the culture chambers is set to a specific number according to actual culture requirements, and each row is guaranteed to be an even number to be matched with the liquid inlet channels 502 of the organoid culture chambers; organoid culture chamber 501 communicates with organoid culture chamber fluid inlet 102 via organoid culture chamber fluid inlet channel 502; the liquid inlet channel 502 of the organoid culture chamber is in a Y-shaped bifurcation structure, and the structure can ensure that liquid is equally divided into the next layer of flow channels each time of bifurcation, so that the liquid injected from the liquid injection port of the organoid culture chamber can evenly flow into each organoid culture chamber 501. The medium channel 401 and the medium reservoir 201 communicate through the medium reservoir outlet 301 provided in the intermediate layer 3; the medium channel 401 and the liquid evaporation chamber 202 communicate through a liquid evaporation chamber liquid inlet 302 provided in the intermediate layer 3.
The main body of the culture medium channel 401 is of a rectangular structure, and two ends of the culture medium channel are of triangular structures, so that a hexagonal structure is formed; the inside of the device is of a cambered surface structure, no sharp angle is contained, and the liquid inlet end and the liquid outlet end are of smooth triangle structures; as shown in fig. 1, the main body has a rectangular structure, and two ends have triangular structures to form a hexagonal structure; one end is communicated with a liquid outlet 301 of the culture medium storage chamber, and the other end is communicated with a liquid inlet 302 of the liquid evaporation chamber. The top layer 1 is provided with a medium storage chamber liquid inlet 101 and a liquid evaporation chamber outlet 103. The width of the culture medium channel 401 is adjusted according to the number and bottom area of the organoid culture chambers 501, so that all the organoid culture chambers 501 can be completely covered.
The organoid culture chambers 501 are provided with two rows, and the organoid culture chamber fluid inlet 102 is provided with two fluid inlets to the organoid culture chambers 501 of the two rows, respectively.
The top layer 1, the storage layer 2, the middle layer 3, the culture medium layer 4, the culture chamber layer 5 and the bottom layer 6 are combined into a whole structure by a vacuum plasma bonding method. The top layer 1, the storage layer 2, the middle layer 3, the culture medium layer 4, the culture chamber layer 5 and the bottom layer 6 are formed by transparent material through mold opening injection molding, and can be prepared by other methods.
The whole chip structure can be formed by adopting transparent material mold opening injection molding, and the transparent material comprises PS, PC, PMMA and can also be made of flexible polydimethylsiloxane materials; of course, other transparent materials may be used. The vacuum plasma bonding mode is adopted to combine into a whole, and the vacuum plasma bonding mode can also combine into a whole through other methods.
The area of the liquid evaporation chamber outlet 103 was 0.32 cm 2 ～4.5 cm 2 The method comprises the steps of carrying out a first treatment on the surface of the The size of the outlet area can be adjusted according to the required perfusion flow rate, and the size of the outlet area is 4.5 cm according to the test result 2 The liquid evaporation rate was 14 to 17.5. Mu.L/h when the area was 2. 2 cm 2 When the liquid evaporation speed is 6-10 mu L/h; when the area size is 1 cm 2 The liquid evaporation rate is 5-8.5 mu L/h, and the area is 0.32-0.32 cm 2 The liquid evaporation rate was 2.5-4.2. Mu.L/h. The specific evaporation speed is different according to different internal air flow circulation structures of different types of incubators. The width of the culture medium channel can be adjusted according to the size of the cell culture chamber, and the depth can be adjusted according to the required liquid perfusion flow rate. The liquid perfusion flow rate of the culture medium channel can be calculated according to the actual width, depth and liquid evaporation speed of the culture medium channel.
The device comprises a culture medium storage chamber, a plurality of organoid culture chambers and a liquid evaporation chamber communicated with the external environment; the medium storage chamber and the liquid evaporation chamber are communicated by a channel to form a communicating vessel. The complete coverage organoid culture chamber of passageway provides nutrient exchange for organoid in the organoid culture chamber. The liquid amount in the evaporating chamber is reduced by the evaporating action of the constant temperature condition of 37 ℃ in the cell incubator on the medium in the evaporating chamber, and the driving force can be provided for the movement of the fluid in the chip under the action of the communicating vessel principle. The liquid in the culture medium storage chamber slowly and continuously flows to the evaporation chamber, and the nutrient substances of the organoid culture in the organoid culture chamber are updated and supplemented by the culture medium flowing through the culture medium channel. The device gets rid of the dependence on liquid driving force equipment completely, can provide driving force for the flow of the culture medium to the inside of the chip by virtue of the liquid self-evaporation effect and the communicating vessel principle, simulates the blood circulation process of continuous natural flow in the internal blood supply environment, and realizes the update and supplement of the culture medium for the organoid in the organoid culture chamber. The control of the liquid flow is realized through the evaporation surface areas with different sizes, the dependence on equipment is eliminated, and the dynamic 3D perfusion culture can be realized by the single-hole organoid chip.
Claims (10)
1. The pump-driven organoid chip is characterized by comprising a top layer (1), a storage layer (2), a middle layer (3), a culture medium layer (4), a culture chamber layer (5) and a bottom layer (6) which are sequentially stacked from top to bottom to form an integral structure;
the storage layer (2) is provided with a culture medium storage chamber (201) and a liquid evaporation chamber (202) communicated with the outside; the surface of the outlet (103) of the liquid evaporation chamber is covered with a microporous filter membrane (7) with the thickness of 0.22 mu m;
the culture chamber layer (5) is provided with a plurality of organoid culture chambers (501), and the organoid culture chambers (501) are communicated with the organoid culture chamber liquid inlet (102) arranged on the top layer (1); the liquid inlet (102) of the organoid culture chamber sequentially passes through the storage layer (2), the middle layer (3) and the culture medium layer (4) to be communicated with the organoid culture chamber (501);
the culture medium layer (4) is provided with a culture medium channel (401), and the culture medium channel (401) is communicated with the liquid evaporation chamber (202), the culture medium storage chamber (201) and the organoid culture chamber (501).
2. The pump-free organoid chip of claim 1, wherein the organoid culture chambers (501) are arranged in an array.
3. The pump-free organoid chip of claim 2, wherein the organoid culture chamber (501) is in communication with the organoid culture chamber fluid inlet (102) via the organoid culture chamber fluid inlet channel (502); the liquid inlet channel (502) of the organoid culture chamber is of a Y-shaped bifurcation structure.
4. A pump-free organoid chip as claimed in claim 3, wherein the organoid culture chambers (501) are provided in two rows and the organoid culture chamber inlet (102) is provided with two inlets for each of the two rows of organoid culture chambers (501).
5. The pump-free organoid chip of claim 1, wherein the medium channel (401) and the medium reservoir (201) are in communication via a medium reservoir outlet (301) provided in the intermediate layer (3); the culture medium channel (401) is communicated with the liquid evaporation chamber (202) through a liquid inlet (302) of the liquid evaporation chamber arranged on the middle layer (3).
6. The pump-free organoid chip of claim 5, wherein said medium channel (401) has a rectangular configuration and triangular configuration at both ends, forming a hexagonal configuration; one end of the liquid evaporator is communicated with a liquid outlet (301) of the culture medium storage chamber, and the other end of the liquid evaporator is communicated with a liquid inlet (302).
7. A pump-free organoid chip according to claim 1, characterized in that the top layer (1) is provided with a medium reservoir inlet (101) and a liquid evaporation chamber outlet (103).
8. The pump-free organoid chip of claim 5, wherein the organoid culture chambers (501) are provided in two rows and the organoid culture chamber fluid inlet (102) is provided with two fluid inlets to the organoid culture chambers (501) in two rows, respectively.
9. The pump-free organoid chip of claim 7, wherein the liquid vaporization chamber outlet (103) has an area of 0.32 cm 2 ～4.5 cm 2 。
10. The pump-free organoid chip of claim 1, wherein the top layer (1), the reservoir layer (2), the middle layer (3), the culture medium layer (4), the culture chamber layer (5) and the bottom layer (6) are made of transparent materials, and are made of one of polydimethylsiloxane, polystyrene plastic and polymethyl methacrylate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311524199.3A CN117229917A (en) | 2023-11-16 | 2023-11-16 | Pump-driving-free organoid chip |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311524199.3A CN117229917A (en) | 2023-11-16 | 2023-11-16 | Pump-driving-free organoid chip |
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| CN117229917A true CN117229917A (en) | 2023-12-15 |
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| CN101817495A (en) * | 2010-03-25 | 2010-09-01 | 湖南大学 | Micro fluid control chip and preparation method and application thereof |
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| CN113801788A (en) * | 2021-08-30 | 2021-12-17 | 西安理工大学 | Cell culture device and method for monitoring cell growth state in real time |
| CN114854583A (en) * | 2022-03-31 | 2022-08-05 | 珠海横琴国草堂中医药科技有限公司 | Cell culture chip and single circulation fluid device |
| WO2023037047A1 (en) * | 2021-09-09 | 2023-03-16 | Finnadvance Oy | Microfluidic cell culture device and method for cell cultivation |
| CN116083234A (en) * | 2023-02-23 | 2023-05-09 | 广州市第一人民医院（广州消化疾病中心、广州医科大学附属市一人民医院、华南理工大学附属第二医院） | Organoid array chip and drug screening method and application thereof |
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-
2023
- 2023-11-16 CN CN202311524199.3A patent/CN117229917A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101817495A (en) * | 2010-03-25 | 2010-09-01 | 湖南大学 | Micro fluid control chip and preparation method and application thereof |
| US20110263008A1 (en) * | 2010-04-26 | 2011-10-27 | National Changhua University Of Education | Cell Culture Real-Time Observation System |
| CN113801788A (en) * | 2021-08-30 | 2021-12-17 | 西安理工大学 | Cell culture device and method for monitoring cell growth state in real time |
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