CN115997732A - Microfluidic nematode culture test system - Google Patents
Microfluidic nematode culture test system Download PDFInfo
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- CN115997732A CN115997732A CN202211635414.2A CN202211635414A CN115997732A CN 115997732 A CN115997732 A CN 115997732A CN 202211635414 A CN202211635414 A CN 202211635414A CN 115997732 A CN115997732 A CN 115997732A
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Abstract
The invention discloses a microfluidic nematode culture test system which comprises a microfluidic control device, a waste liquid collecting device, a shooting device and a control device. The microfluidic control device comprises a microfluidic chip, a liquid pump and a liquid storage container, the microfluidic chip is provided with a plurality of stimulation culture cavities which are arranged in parallel, each stimulation culture cavity is provided with a liquid inlet, a liquid outlet and a nematode loading inlet, the liquid pump is used for pumping liquid towards the liquid storage container and pressing the liquid in the liquid storage container into the stimulation culture cavity, the waste liquid collecting device is communicated with the liquid outlet to collect waste liquid, and the shooting device is used for shooting and storing the images of the insects in the stimulation culture cavities. The control device is electrically connected with the micro-flow control device and the shooting device. The microfluidic nematode culture test system can effectively solve the problems of inaccurate solution concentration control and poor uniformity of different batches in the existing nematode chemical toxicity evaluation test process, improves the experimental efficiency, and facilitates systematic research of nematode toxicity tests.
Description
Technical Field
The invention relates to the technical field of biological experimental equipment, in particular to a microfluidic nematode culture test system.
Background
In recent years, nematodes have demonstrated unique advantages in chemical toxicity evaluation tests. Compared with other model organisms, the nematode test period is short, the breeding quantity is large, the nematode test period is small, ethical examination is not needed, the nematode test period is easy to culture, the nematode test period is low in cost, transparent throughout bodies are easy to observe cells and organs with high precision, and the nematode test period has complete sequencing of genome library and gene plasticity, so that toxicity evaluation test and analysis are greatly facilitated. However, the existing nematode toxicity test is still manually completed by an experimenter, and further systematic research on nematode toxicity test is seriously hindered by the problems of complicated operation steps, inaccurate concentration control (agar absorption), insufficient uniformity of different batches, low throughput, low experimental efficiency and the like by adopting a porous plate or an agar culture dish.
Disclosure of Invention
The invention aims to provide a microfluidic nematode culture test system which can effectively solve the problems of inaccurate solution concentration control and poor uniformity of different batches in the existing nematode chemical toxicity evaluation test process, improves the experimental efficiency and facilitates systematic research of nematode toxicity tests.
In order to achieve the technical effects, the technical scheme of the invention is as follows:
the invention discloses a microfluidic nematode culture test system, which comprises: a microfluidic control device comprising a microfluidic chip having a plurality of stimulation culture chambers arranged in parallel, each of the stimulation culture chambers having a liquid inlet, a liquid outlet and a nematode loading inlet, a liquid pump for pumping liquid towards the liquid storage container and pressing the liquid within the liquid storage container into the stimulation culture chambers; a waste liquid collection device in communication with the liquid outlet to collect waste liquid; the shooting device is used for shooting and storing the line worm images in the stimulation culture cavity; and the control device is electrically connected with the micro-flow control device and the shooting device.
In some embodiments, the liquid storage container comprises a nematode culture liquid bottle, a nematode food suspension bottle, and a chemical bottle to be tested, disposed in parallel.
In some specific embodiments, the number of chemical bottles to be tested is at least two.
In some embodiments, the photographing device includes a microscope located above the microfluidic chip and a CCD camera for photographing an image of the microscope.
In some embodiments, the microfluidic nematode culture test system further comprises a moving platform for supporting the microfluidic chip, the moving platform for driving the microfluidic chip to move along a first direction and a second direction, the first direction and the second direction being vertically arranged.
In some specific embodiments, the mobile platform comprises: the bearing platform is used for supporting the microfluidic chip; the movable end of the first driving module is connected with the bearing platform so as to drive the bearing platform to move along the first direction; the movable end of the second driving module is connected with the fixed end of the first driving module so as to drive the first driving module and the bearing platform to move along the second direction.
In some embodiments, the microfluidic nematode culture test system further comprises an egg analysis device coupled to the waste collection device, the egg analysis device capturing and immobilizing eggs in the waste and incubating.
In some specific embodiments, the egg analysis device includes a liquid flow channel and a plurality of egg capturing culture chambers connected to the liquid flow channel, wherein the plurality of egg capturing culture chambers are arranged in rows and columns, two adjacent rows of egg capturing culture chambers are arranged in parallel, and the plurality of egg capturing culture chambers in the same row are arranged in series.
In some more specific embodiments, each of the egg capturing culture chambers accommodates one egg, and each of the egg capturing culture chambers is connected in parallel with one bypass flow path.
In some specific embodiments, the liquid flow channel has a nematode culture liquid inlet, a food suspension liquid inlet, a waste liquid inlet, and a waste liquid outlet, the waste liquid inlet is connected to the waste liquid collection device, and the nematode culture liquid inlet, the food suspension liquid inlet, and the waste liquid inlet are in communication with the inlet of the egg capturing culture chamber, and the waste liquid outlet is in communication with the outlet of the egg capturing culture chamber.
The microfluidic nematode culture test system provided by the embodiment of the invention has the beneficial effects that: the microfluidic nematode culture test system of the embodiment is adopted, a porous plate or an agar culture dish is not needed to be manually adopted for nematode toxicity test, the problems that the solution concentration control is inaccurate and the uniformity is poor in different batches in the existing nematode chemical toxicity evaluation test process are effectively solved, the experimental efficiency is improved, and the systematic research of nematode toxicity test is facilitated.
Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.
Drawings
FIG. 1 is a schematic diagram of a microfluidic nematode culture test system according to an embodiment of the present invention;
fig. 2 is a schematic structural diagram of a microfluidic chip according to an embodiment of the present invention;
FIG. 3 is a schematic diagram showing the structure of an insect egg analyzing apparatus according to an embodiment of the present invention;
FIG. 4 is a schematic view showing a partial structure of an egg analyzing apparatus according to an embodiment of the present invention.
Reference numerals:
1. a control device; 2. a CCD camera; 3. a microscope; 4. a microfluidic chip; 41. stimulating the culture cavity; 411. a liquid inlet; 412. a liquid outlet; 413. a nematode loading inlet; 42. a liquid inlet flow channel; 43. a liquid outlet channel; 5. a liquid pump; 6. a nematode culture solution bottle; 7. a nematode food suspension bottle; 8. a chemical bottle to be tested; 9. a waste liquid collection device; 10. a mobile platform; 20. an insect egg analysis device; 201. a liquid flow channel; 202. an insect egg capturing culture cavity; 203. a branch flow passage; 204. a nematode culture solution inlet; 205. a food suspension inlet; 206. a waste liquid inlet; 207. and a waste liquid outlet.
Detailed Description
In order to make the technical problems solved, the technical scheme adopted and the technical effects achieved by the invention more clear, the technical scheme of the invention is further described below by a specific embodiment in combination with the attached drawings.
In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the device or element being referred to must have a specific orientation, be configured and operated in a specific orientation, and therefore should not be construed as limiting the present invention.
Furthermore, features defining "first", "second" may include one or more such features, either explicitly or implicitly, for distinguishing between the descriptive features, and not sequentially, and not lightly. In the description of the present invention, unless otherwise indicated, the meaning of "a plurality" is two or more.
In the description of the present invention, it should be noted that, unless explicitly specified and limited otherwise, the terms "mounted," "connected," and "connected" are to be construed broadly, and may be either fixedly connected, detachably connected, or integrally connected, for example; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present invention will be understood in specific cases by those of ordinary skill in the art.
The specific structure of the microfluidic nematode culture test system according to the embodiment of the present invention is described below with reference to fig. 1 to 4.
The invention discloses a microfluidic nematode culture test system, which is shown in fig. 1-2, and comprises a microfluidic control device, a waste liquid collecting device 9 and a shooting device, wherein the microfluidic control device comprises a microfluidic chip 4, a liquid pump 5 and a liquid storage container, the microfluidic chip 4 is provided with a plurality of stimulation culture cavities 41 which are arranged in parallel, each stimulation culture cavity 41 is provided with a liquid inlet 411, a liquid outlet 412 and a nematode loading inlet 413, the liquid pump 5 is used for pumping liquid towards the liquid storage container and pressing the liquid in the liquid storage container into the stimulation culture cavities 41, the waste liquid collecting device 9 is communicated with the liquid outlet 412 to collect waste liquid, the shooting device is used for shooting and storing the images of the nematodes in the stimulation culture cavities 41, and the control device 1 is electrically connected with the microfluidic control device and the shooting device.
It will be appreciated that the control device 1 controls the liquid pump 5 in the microfluidic control device to pump (flow-controllable) the solution in the liquid storage container continuously into the microfluidic chip 4, and the liquid is fed into the stimulation culture chamber 41 through the distribution flow channel on the microfluidic chip 4, the solution flow in the chamber is kept fresh, and then individual nematodes are transferred into the nematode loading inlet 413 of the microfluidic chip 4 using the nematode pick. Therefore, in the actual working process, parameters such as solutions of different types, solutions of different concentrations, exposure time of each solution and the like can be controlled by programming control instructions of the control device 1, so that free combination is realized. In the experimental process, the shooting device is used for shooting the nematode image in the stimulation culture cavity 41, storing the image and feeding back the related information into the control device 1, and the control device 1 contains data analysis software to perform real-time online analysis on the image data so as to extract the commonly used nematode toxicity test evaluation index. The nematode morphological characteristics including length, width and movement characteristics including swimming amplitude, frequency and the like can be analyzed online in real time.
In summary, in the experimental process, the liquid flow can be precisely controlled by the microfluidic chip 4, and the shooting device shoots and stimulates the nematode image in the culture cavity 41 and stores and feeds back the relevant information to the control device 1, so that real-time online analysis is realized.
In the present embodiment, the control device 1 is a host computer of a high content information analysis program for computer vision, and the related program is written based on OpenCV computer vision technology. The high content information analysis program includes the following information: the nematode exercise behavior, individual development characteristics, reproduction information, mortality and life cycle, and other customized evaluation indexes are subjected to information extraction and analysis. Of course, in other embodiments of the present invention, the specific type of the control device 1 may also be defined according to actual needs.
In some embodiments, as shown in fig. 1, the liquid storage container includes a nematode culture liquid bottle 6, a nematode food suspension bottle 7, and a chemical bottle 8 to be tested, which are disposed in parallel. It will be appreciated that the nematode culture liquid bottle 6 and the nematode food suspension liquid bottle 7 can provide a suitable living environment for nematodes in the stimulating culture chamber 41, so as to avoid the phenomenon that nematodes die due to starvation or insufficient nutrition during toxicity test, and the chemical bottle 8 to be tested can be used for placing chemical solution according to actual needs, so as to meet the requirement of toxicity test.
In some embodiments, as shown in FIG. 1, there are at least two bottles 8 of chemical to be tested. Of course, in other embodiments of the invention, the number of chemical bottles 8 to be tested may be selected according to actual needs.
In some embodiments, as shown in fig. 1, the photographing device includes a microscope 3 and a CCD camera 2, the microscope 3 is located above the microfluidic chip 4, and the CCD camera 2 is used to photograph an image of the microscope 3. It can be understood that a clearer image can be obtained by amplifying the microscope 3 and shooting the image by the CCD camera 2, so that the control device 1 can conveniently perform image analysis. Of course, in other embodiments of the present invention, the photographing device may be selected according to actual needs.
In some embodiments, as shown in fig. 1, the microfluidic nematode culture test system further includes a moving platform 10, where the moving platform 10 is used to support the microfluidic chip 4, and the moving platform 10 is used to drive the microfluidic chip 4 to move along a first direction and a second direction, and the first direction and the second direction are perpendicular to each other. It will be appreciated that, according to the foregoing, the microfluidic chip 4 has a plurality of stimulus incubation cavities 41 arranged in parallel, and in the experimental process, the photographing device needs to photograph a plurality of stimulus incubation cavities 41 in sequence, and the microfluidic chip 4 is arranged on the moving platform 10, and in the actual photographing process, the microfluidic chip 4 can be moved under the moving platform 10 to the position below the photographing device, so as to realize automatic photographing of a plurality of stimulus incubation cavities 41.
In some specific embodiments, the mobile platform 10 includes a carrying platform, a first driving module and a second driving module, the carrying platform is used for supporting the microfluidic chip 4, the movable end of the first driving module is connected with the carrying platform to drive the carrying platform to move along the first direction, and the movable end of the second driving module is connected with the fixed end of the first driving module to drive the first driving module and the carrying platform to move along the second direction. It can be understood that the first driving module and the second driving module realize the bidirectional driving of the bearing platform, so that the structure of the mobile platform 10 can be simplified while the micro-fluidic chip 4 is ensured to stably move below the shooting device, and the manufacturing cost of the micro-fluidic nematode culture test system is reduced. It should be noted that, in this embodiment, the first driving module and the second driving module may select any structure such as a motor driving roller, a cylinder, and an electric push rod according to actual needs.
In some embodiments, as shown in fig. 3, the microfluidic nematode culture test system further comprises an egg analysis device 20, wherein the egg analysis device 20 is connected to the waste liquid collection device 9, and the egg analysis device 20 captures and immobilizes eggs in the waste liquid and incubates. It will be appreciated that the worm egg analysis apparatus 20 is capable of facilitating observation and extraction of nematodes and analysis of various evaluation indicators of nematode offspring, thereby expanding the range of application of microfluidic nematode culture test systems.
In some specific embodiments, as shown in fig. 3, the egg analysis device 20 includes a liquid flow channel 201 and a plurality of egg capturing culture chambers 202 connected to the liquid flow channel 201, wherein the plurality of egg capturing culture chambers 202 are arranged in rows and columns, two adjacent rows of egg capturing culture chambers 202 are arranged in parallel, and the plurality of egg capturing culture chambers 202 of the same row are arranged in series. It can be appreciated that the arrangement of a plurality of egg capturing culture chambers 202 enables a plurality of egg rearing at a time, thereby improving experimental efficiency.
In some more specific embodiments, as shown in FIG. 4, each egg capturing culture chamber 202 contains one egg, and each egg capturing culture chamber 202 is connected in parallel with one bypass flow channel 203. It will be appreciated that during actual operation, as each egg capturing culture chamber 202 contains an egg, a single egg, once it enters the egg capturing culture chamber 202, blocks the flow channel and subsequent eggs flow through the lower bypass flow channel 203 to continue screening until all egg capturing culture chambers 202 are filled. Thus, the occurrence of a phenomenon in which a plurality of eggs are accommodated in one egg capturing culture chamber 202, thereby making detection inconvenient, is avoided.
In some embodiments, the liquid flow channel 201 has a nematode culture liquid inlet 204, a food suspension inlet 205, a waste liquid inlet 206, and a waste liquid outlet 207, the waste liquid inlet 206 is connected to the waste liquid collection device 9, and the nematode culture liquid inlet 204, the food suspension inlet 205, and the waste liquid inlet 206 are in communication with the inlet of the egg capturing culture chamber 202, and the waste liquid outlet 207 is in communication with the outlet of the egg capturing culture chamber 202. Thus, it is possible to ensure that the eggs have sufficient nutrient cultivation and that the nematodes have sufficient food growth, thereby facilitating the test.
Examples:
as shown in fig. 1 to 4, the microfluidic nematode culture test system comprises a microfluidic control device, a waste liquid collecting device 9, a photographing device, a mobile platform 10 and an egg analyzing device 20. The micro-flow control device comprises a micro-flow control chip 4, a liquid pump 5 and a liquid storage container, wherein the micro-flow control chip 4 is provided with sixteen stimulation culture cavities 41, a liquid inlet runner 42 and a liquid outlet runner 43 which are arranged in parallel, each stimulation culture cavity 41 is provided with a liquid inlet 411, a liquid outlet 412 and a nematode loading inlet 413, the liquid pump 5 is connected with the liquid storage container, liquid in the liquid storage container is pressed into the stimulation culture cavity 41, and the liquid storage container comprises a nematode culture liquid bottle 6, a nematode food suspension liquid bottle 7 and a chemical bottle 8 to be tested which are arranged in parallel. The liquid storage container is communicated with the liquid inlet 411 through the liquid inlet flow channel 42, the waste liquid collecting device 9 is communicated with the liquid outlet 412 through the liquid outlet flow channel 43 to collect waste liquid, the shooting device comprises a microscope 3 and a CCD camera 2, the microscope 3 is located above the microfluidic chip 4, the CCD camera 2 is used for shooting an image of the microscope 3, and the control device 1 is electrically connected with the microfluidic control device and the shooting device. The mobile platform 10 is used for supporting the microfluidic chip 4, the mobile platform 10 comprises a bearing platform, a first driving module and a second driving module, the bearing platform is used for supporting the microfluidic chip 4, the movable end of the first driving module is connected with the bearing platform to drive the bearing platform to move along a first direction, and the movable end of the second driving module is connected with the fixed end of the first driving module to drive the first driving module and the bearing platform to move along a second direction. The egg analysis device 20 includes a liquid flow channel 201 and a plurality of egg capturing culture chambers 202 connected to the liquid flow channel 201, wherein forty eight egg capturing culture chambers 202 are arranged in six rows and eight columns, two adjacent rows of egg capturing culture chambers 202 are arranged in parallel, and a plurality of egg capturing culture chambers 202 in the same row are arranged in series. Each egg capturing culture chamber 202 accommodates one egg, and each egg capturing culture chamber 202 is connected in parallel with one bypass flow passage 203. The liquid flow channel 201 has a nematode culture liquid inlet 204, a food suspension liquid inlet 205, a waste liquid inlet 206, and a waste liquid outlet 207, the waste liquid inlet 206 is connected to the waste liquid collecting device 9, and the nematode culture liquid inlet 204, the food suspension liquid inlet 205, and the waste liquid inlet 206 are communicated with an inlet of the egg capturing culture chamber 202, and the waste liquid outlet 207 is communicated with an outlet of the egg capturing culture chamber 202.
In the description of the present specification, reference to the term "some embodiments," "other embodiments," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The foregoing is merely exemplary of the present invention, and those skilled in the art should not be considered as limiting the invention, since modifications may be made in the specific embodiments and application scope of the invention in light of the teachings of the present invention.
Claims (10)
1. A microfluidic nematode culture test system, comprising:
a microfluidic control device comprising a microfluidic chip (4), a liquid pump (5) and a liquid storage container, the microfluidic chip (4) having a plurality of stimulation culture chambers (41) arranged in parallel, each stimulation culture chamber (41) having a liquid inlet (411), a liquid outlet (412) and a nematode loading inlet (413), the liquid pump (5) being for pumping liquid towards the liquid storage container and for pressing liquid within the liquid storage container into the stimulation culture chambers (41);
a waste liquid collection device (9), the waste liquid collection device (9) being in communication with the liquid outlet (412) to collect waste liquid;
the shooting device is used for shooting and storing the wire worm images in the stimulation culture cavity (41);
and a control device (1), wherein the control device (1) is electrically connected with the micro-flow control device and the shooting device.
2. Microfluidic nematode culture test system according to claim 1, characterized in that the liquid storage container comprises a nematode culture liquid bottle (6), a nematode food suspension bottle (7) and a chemical bottle to be tested (8) arranged in parallel.
3. Microfluidic nematode culture test system according to claim 2, characterized in that the number of chemical bottles (8) to be tested is at least two.
4. The microfluidic nematode culture test system according to claim 1, wherein the photographing device comprises a microscope (3) and a CCD camera (2), the microscope (3) is located above the microfluidic chip (4), and the CCD camera (2) is used for photographing imaging of the microscope (3).
5. The microfluidic nematode culture test system according to claim 1, further comprising a moving platform (10), the moving platform (10) being configured to support the microfluidic chip (4), the moving platform (10) being configured to drive the microfluidic chip (4) to move in a first direction and a second direction, the first direction and the second direction being arranged perpendicularly.
6. The microfluidic nematode culture test system according to claim 5, wherein the mobile platform (10) comprises:
the bearing platform is used for supporting the microfluidic chip (4);
the movable end of the first driving module is connected with the bearing platform so as to drive the bearing platform to move along the first direction;
the movable end of the second driving module is connected with the fixed end of the first driving module so as to drive the first driving module and the bearing platform to move along the second direction.
7. Microfluidic nematode culture test system according to any of claims 1-6, characterized in that it further comprises an egg analysis device (20), which egg analysis device (20) is connected to the waste liquid collection device (9), which egg analysis device (20) captures and immobilizes eggs in the waste liquid and incubates.
8. The microfluidic nematode culture test system according to claim 7, wherein the worm egg analysis device (20) comprises a liquid flow channel (201) and a plurality of worm egg capturing culture chambers (202) connected to the liquid flow channel (201), the worm egg capturing culture chambers (202) are arranged in rows and columns, two adjacent worm egg capturing culture chambers (202) are arranged in parallel, and the worm egg capturing culture chambers (202) of the same row are arranged in series.
9. The microfluidic nematode culture test system according to claim 8, wherein each egg capturing culture chamber (202) accommodates one egg, and each egg capturing culture chamber (202) is connected in parallel with a bypass flow channel (203).
10. The microfluidic nematode culture test system according to claim 8, wherein the liquid flow channel (201) has a nematode culture liquid inlet (204), a food suspension inlet (205), a waste liquid inlet (206) and a waste liquid outlet (207), the waste liquid inlet (206) is connected to the waste liquid collecting device (9), and the nematode culture liquid inlet (204), the food suspension inlet (205) and the waste liquid inlet (206) are in communication with the inlet of the egg capturing culture chamber (202), and the waste liquid outlet (207) is in communication with the outlet of the egg capturing culture chamber (202).
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