CN115138402A - Micro-fluidic chip capable of setting chemical concentration gradient and preparation method and application thereof - Google Patents
Micro-fluidic chip capable of setting chemical concentration gradient and preparation method and application thereof Download PDFInfo
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/06—Quantitative determination
Abstract
The invention relates to a micro-fluidic chip capable of setting chemical concentration gradient and a preparation method and application thereof. Specifically, the method comprises the following steps: 1) Obtaining a microfluidic chip with a channel with a specific size, wherein the channel on the microfluidic chip comprises a concentration gradient forming part and sample injection parts arranged at two ends of the concentration gradient forming part, and the depth of the channel on the microfluidic chip is less than 80 mu m; 2) Preparing a melted agar gel solution, wherein the concentration of the agar gel solution is 0.15-0.3%; 3) And injecting the agar gel solution into a channel of the microfluidic chip, and standing to form the agar gel capable of forming a chemical concentration gradient. The channel of the invention has simple design, the gradient is stably kept for more than 12 hours, and the concentration gradient difference of 5-100% can be realized by adjusting the size of the channel.
Description
Technical Field
The invention belongs to the field of microfluidics, and particularly relates to a chemical concentration gradient establishment method based on a microfluidic chip.
Background
The microfluidic technology refers to a set of technologies for operating micro-volume (mL, nL, pL) liquid in a manually manufactured microsystem, has the capability of scaling down the basic functions of laboratories such as biology, chemistry and the like, such as sample preparation, reaction, separation, detection and the like, to a chip with the square centimeter, and has the basic characteristics and the greatest advantage that various unit technologies are flexibly combined and integrated on a whole controllable micro platform in a large scale.
Biochemical molecules such as growth factors, protein signals, chemoattractants, etc. often form chemical concentration gradients that play an important role in many biological processes. Especially, microorganisms often face complicated and variable environments in nature, so that many microorganisms develop chemical concentrations in environments with efficient chemotactic systems, and move along concentration gradients to achieve the purpose of benefiting and avoiding harm. To study these complex biological phenomena, scientists need to establish a concentration gradient of chemical substances in vitro to mimic the microenvironment in which the organism is located. The existing microfluidic technology for establishing chemical concentration gradients is to use laminar flow between liquid flows to transfer mass between parallel liquid flows by diffusion, forming a concentration gradient perpendicular to the direction of the liquid flows. By controlling the liquid flow rate and the contact time, a linear concentration gradient can be formed quickly and can be kept stable for a long time under the condition of continuously providing liquid flow. The common micro-fluidic chip structure comprises a T-shaped structure and a Christmas tree structure, wherein the T-shaped structure is formed by connecting two or more micro channels to one micro channel, the Christmas tree structure is formed by designing a plurality of layers of micro channels which are constantly branched to realize repeated flow distribution and convergence, and finally, a mixed concentration gradient is formed in one main channel.
Types of conventional concentration gradient generators include Zigmomd chambers, boyden chemotactic chambers, dunn chambers, bio-gels, and the like. However, these conventional methods are difficult to achieve precise and controllable adjustment of the chemical concentration gradient and maintain for a short period of time. Compared with the traditional method, the microfluidic technology has the advantages that a stable, controllable and quantifiable chemical substance concentration gradient can be formed, the structure of the microchannel can be designed and processed according to experimental requirements, and the movement track of the single cell can be tracked more favorably by combining the microscope technology.
The most common concentration gradient generator based on the microfluidic technology at present is a parallel flow generator, and the technology has the disadvantages that firstly, a liquid flow needs to be continuously provided to maintain the stability of the concentration gradient, so that a large amount of reagents are consumed; second, the continuous flow of liquid can cause mechanical forces to damage cells or bacteria; third, complex structural designs are often required to achieve precise control of concentration. Therefore, the prior art lacks a concentration gradient microfluidic chip which has a simple structure, can quickly form a stable concentration gradient and does not need to consume a large amount of expensive reagents.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention designs the micro-fluidic chip with a specific size, can generate a stable concentration gradient by only using agar gel with a certain concentration to control the diffusion rate of a compound, keeps the stability for a long time, and simultaneously reduces the damage of the shearing force of a liquid flow to cells.
One aspect of the present invention provides a method for preparing a microfluidic chip capable of setting a chemical concentration gradient, the method comprising the steps of:
1) Obtaining a micro-fluidic chip with a channel with a specific size, wherein the channel on the micro-fluidic chip comprises a concentration gradient forming part and sample injection parts arranged at two ends of the concentration gradient forming part, and the depth of the channel on the micro-fluidic chip is less than 80 mu m; the channel on the microfluidic chip is punched to the channel on the PDMS layer at the position of the sample injection part through the PDMS layer with the groove of the concentration gradient forming part and the sample injection part to form a sample injection hole, and then the sample injection hole is bonded with the glass slide to form a sealed microfluidic chip;
2) Preparing an agar gel solution, wherein the concentration of the agar solution is 0.15% -0.3%;
3) And injecting the agar solution into a channel of the microfluidic chip, and standing to form agar gel capable of forming chemical concentration gradient.
In the technical scheme of the invention, the method further comprises the step 4) of injecting the active ingredients andor the dye into the micro-fluidic chip through the sampling hole at one side of the channel, and standing to obtain the micro-fluidic chip with the concentration gradient.
In the technical scheme of the invention, the cross section of the channel on the microfluidic chip is rectangular.
In the technical scheme of the invention, the depth of the channel on the microfluidic chip is 40-55 μm.
In the technical scheme of the invention, the width of the channel on the microfluidic chip is 0.8-2.5mm.
In the technical scheme of the invention, the length of the channel on the microfluidic chip is 0.8-2.5cm.
In the technical scheme of the invention, the aperture of the sampling hole is 50-100% of the width of the channel on the microfluidic chip, and preferably 1.1-1.5mm.
In the technical scheme of the invention, the concentration of the agar gel solution is 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29% and 0.30%.
In the technical scheme of the invention, the number of the channels on the microfluidic chip is more than one.
The invention provides another microfluidic chip capable of setting chemical concentration gradient, wherein a channel is arranged on the microfluidic chip and comprises a concentration gradient forming part and sample injection parts arranged at two ends of the concentration gradient forming part, and the depth of the channel on the microfluidic chip is less than 80 μm; the channel on the microfluidic chip is punched to the channel on the PDMS layer at the position of the sample injection part through the PDMS layer with the groove of the concentration gradient forming part and the sample injection part to form a sample injection hole, and then the sample injection hole is bonded with the glass slide to form a sealed channel containing the concentration gradient forming part and the sample injection part; and the channel also comprises agar gel, and the concentration of the agar gel is 0.15% -0.3%.
In the technical scheme of the invention, the cross section of the channel on the microfluidic chip is rectangular.
In the technical scheme of the invention, the depth of the channel on the microfluidic chip is 40-55 μm.
In the technical scheme of the invention, the width of the channel on the microfluidic chip is 0.8-2.5mm.
In the technical scheme of the invention, the length of the channel on the microfluidic chip is 0.8-2.5cm.
In the technical scheme of the invention, the aperture of the sample inlet hole is 50-100% of the width of the channel on the microfluidic chip.
In the technical scheme of the invention, the number of the channels on the microfluidic chip is more than one.
In the technical scheme of the invention, the concentration gradient forming part of the channel on the microfluidic chip is a strip-shaped groove with a rectangular cross section, and the sample inlet part and the sample outlet part are respectively communicated with the concentration gradient forming part.
In a further aspect, the invention provides an application of the microfluidic chip, wherein the application is a tracking device for microorganisms or cells under a low power lens.
In a further aspect of the invention, there is provided a microorganism or cell tracking device comprising a microfluidic chip according to the invention as described above and an observation or recording device, preferably selected from a fluorescence inverted microscope.
Advantageous effects
The invention has the advantages that the channel design is simple, the liquid flow is not required to be continuously provided, different concentration gradients can be generated by controlling the diffusion rate of the chemical by using agar with different concentrations, and the stable maintenance can be carried out for 12 hours. Since bacteria or cells are not directly exposed to the liquid stream, damage from shear forces is reduced. Can ensure that the concentration gradient of the cells keeps stable in the process of carrying out three-dimensional motion tracking in the chip microchannel. The micro-fluidic chip has simple design and convenient manufacture, and only needs a small amount of reagents; and by adjusting the size of the channel, the concentration gradient difference of 5-100% can be realized.
Drawings
Fig. 1 is a schematic diagram of a channel structure in a microfluidic chip according to the present invention. Wherein, 1 is a concentration gradient forming part, 2 is a sample injection part, and 3 is a sample injection hole. The invention has the advantages that the channel design is simple, the continuous liquid flow is not required to be provided, different concentration gradients can be generated by controlling the diffusion rate of the chemical substances by using the agar with different concentrations, and the stable maintenance can be carried out for 12 hours. Since bacteria or cells are not directly exposed to the liquid stream, damage due to shear forces is reduced. Can ensure that the concentration gradient of the cells keeps stable in the process of carrying out three-dimensional motion tracking in the chip microchannel. The micro-fluidic chip has simple design and convenient manufacture, and only needs a small amount of reagent. By adjusting the size of the channels, a concentration gradient difference of 5-100% can be achieved.
FIG. 2 is a grey scale value analysis of the concentration gradient formed by the fluorescent dye in the channel. The chemical gradient is traced and displayed by using the fluorescent dye, and the fluorescence intensity in the channel can be seen to be reduced from one end of the fluorescent dye injection to the other end in sequence under a microscope. The grey values were used to analyze the distribution data of the fluorochromes to obtain the concentration change of the fluorochromes in the channel, and fig. 3 shows the concentration gradient of the fluorochromes with good performance.
Fig. 3 is a photograph of a microfluidic chip of the present invention, wherein 8 channels are disposed on the microfluidic chip.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, specific embodiments thereof are described in detail below, but the present invention is not to be construed as being limited to the implementable range thereof.
The scheme of the invention is explained in the following with the attached drawings.
The specific embodiment of the invention provides a preparation method of a microfluidic chip capable of setting chemical concentration gradient, which comprises the following steps:
1) Obtaining a micro-fluidic chip with a channel with a specific size, wherein the channel on the micro-fluidic chip comprises a concentration gradient forming part and sample injection parts arranged at two ends of the concentration gradient forming part, and the depth of the channel on the micro-fluidic chip is less than 80 mu m; (ii) a The channel on the microfluidic chip is punched to the channel through the PDMS layer with the groove of the concentration gradient forming part and the sample injection part to form a sample injection hole, and then the sample injection hole is bonded with the glass slide to form the channel comprising the concentration gradient forming part and the sample injection part.
2) Preparing an agar solution, wherein the concentration of the agar solution is 0.15% -0.3%;
3) Injecting the agar solution into the channel of the microfluidic chip, standing to form agar gel capable of forming chemical concentration gradient
In some embodiments of the present invention, the method further comprises step 4) injecting the active ingredient and/or the dye through the sample hole on one side of the channel, and obtaining the microfluidic chip with a concentration gradient after standing.
In some embodiments of the invention, wherein the cross-section of the channel on the microfluidic chip is rectangular.
In some embodiments of the present invention, the depth of the channel on the microfluidic chip is 40-55 μm. In the present invention, the inventors found that the depth of the channel affects the state of the agar gel, and when the depth of the channel is too deep, for example, more than 80 μm, a stable gel structure cannot be formed, and thus an effective concentration gradient cannot be formed; and when the depth is less than 80 μm, a stable gel structure can be formed, and an effective concentration gradient can be formed.
In some embodiments of the invention, the width of the channel on the microfluidic chip is 0.8-2.5mm.
In some embodiments of the invention, the length of the channel on the microfluidic chip is 0.8-2.5cm. In the present invention, since the concentration of the agar gel is low, if the time for forming the concentration gradient is too long, an effective concentration gradient cannot be formed due to the change in the concentration of the agar gel, and therefore, it is necessary to control the length of the channel to be 0.8 to 2.5cm.
In some embodiments of the present invention, the aperture of the sample inlet is 50% to 100%, preferably 1 to 1.5mm, of the width of the channel on the microfluidic chip.
In some embodiments of the invention, the concentration of the agar gel solution is 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.30%. In the invention, the concentration of the agar gel solution is very dilute, so that a uniform and stable gel structure can be formed by matching with a special channel size, and a concentration gradient can be formed after the dye or the active component is added. If the agar concentration is further small, a stable gel structure cannot be formed, while if the concentration is too large, too long a time is required for establishing a gradient, and too long a time results in excessive loss of water in the gel structure to change the gel concentration again.
In some embodiments of the invention, the number of channels on the microfluidic chip is more than one.
In some embodiments of the invention, the microorganisms or single cells are added to the ipsilateral or ipsilateral wells to which the active ingredient or dye is added after the gel is formed and the activity of the microorganisms or single cells can be observed.
In some embodiments of the present invention, in step 1), a microfluidic chip with a channel is obtained by using a photolithography technique, and after the designed channel is drawn into a CAD file, a film with structural information is manufactured by a third party company, which is the same as a conventional process for manufacturing the microfluidic chip. Etching the structure on the film on a silicon wafer through a photoetching process, selecting proper photoetching parameters according to the photoresist properties by using SU8-3050 photoresist, and manufacturing the micro-channel with the height of 40-55 mu m. Finally, the PDMS is flatly laid on a silicon chip with a structure, and the silicon chip is placed in an oven at 80 ℃ to be dried until solidification. And after the PDMS is solidified, the PDMS block is taken off from the silicon chip, the channel structure is cut, a hole is punched at the position of the sample injection part, a sample injection hole penetrating through the upper layer PDMS to reach the channel is formed, and then the PDMS structure is bonded on the clean glass slide through plasma bonding.
The resulting PDMS structure-loaded chip was used to construct chemical concentration gradients. Injecting heated agar gel solution into the sample inlet at one end of the channel, cooling and solidifying after the whole chamber is filled with the heated agar gel solution, and then injecting fluorescent dye from another sample inlet. At this time, the size of the chemical gradient and the time required for the gradient formation can be controlled by adjusting the agar gel concentration, which is set at 0.15% to 0.3%. After a certain time, the concentration gradient is established, then the bacteria sample to be researched is injected into the inlet of the agar gel solution, and the chip can be placed under a microscope for real-time observation after the bacteria generate chemotactic motion in the channel.
In some embodiments of the present invention, the dye is any dye that develops color under natural light or ultraviolet light, such as a fluorescent dye.
In some embodiments of the invention, the active substance includes any substance that acts on a cell or microorganism, such as nutrients, drugs, irritants, and the like.
EXAMPLE 1 preparation of microfluidic chip
1) Obtaining a micro-fluidic chip with a channel with a specific size, wherein the channel on the micro-fluidic chip comprises a concentration gradient forming part and sample injection parts arranged at two ends of the concentration gradient forming part, and the depth of the channel on the micro-fluidic chip is 50 mu m; the width is 1.5mm, and the length is 1cm; preparing a PDMS layer with a groove of a concentration gradient forming part and a sample injection part by adopting a photoetching method, and punching a sample injection hole reaching a channel at the position of the sample injection part of the PDMS layer, wherein the diameter of the sample injection hole is 1.1mm.
2) Preparing a melted agar gel solution, wherein the concentration of the agar solution is 0.15%;
3) And injecting the agar solution into a channel of the microfluidic chip, and standing to form agar gel capable of forming chemical concentration gradient.
Example 2 preparation of microfluidic chip
1) Obtaining a microfluidic chip with a channel with a specific size, wherein the channel on the microfluidic chip comprises a concentration gradient forming part and sample injection parts arranged at two ends of the concentration gradient forming part, and the depth of the channel on the microfluidic chip is 45 mu m; the width is 1.5mm, and the length is 1cm; preparing a PDMS layer with a groove of a concentration gradient forming part and a sample injection part by adopting a photoetching method, punching a sample injection hole reaching a channel at the position of the sample injection part of the PDMS layer, wherein the diameter of the sample injection hole is 1.1mm, and then bonding with a glass slide.
2) Preparing a melted agar gel solution, wherein the concentration of the agar solution is 0.15%;
3) And injecting the agar gel solution into a channel of the microfluidic chip, and standing to form the agar gel capable of forming a chemical concentration gradient.
Example 3 preparation of microfluidic chip
1) Obtaining a micro-fluidic chip with a channel with a specific size, wherein the channel on the micro-fluidic chip comprises a concentration gradient forming part and sample injection parts arranged at two ends of the concentration gradient forming part, and the channel on the micro-fluidic chip has the depth of 80 mu m, the width of 1.5mm and the length of 1cm; preparing a PDMS layer with a concentration gradient forming part and a groove of a sample injection part by adopting a photoetching method, then punching a sample injection hole reaching a channel at the position of the sample injection part of the PDMS layer, wherein the diameter of the sample injection hole is 1.1mm, and then bonding with a glass slide;
2) Preparing an agar gel solution, wherein the concentration of the agar gel solution is 0.15%;
3) And injecting the agar gel solution into a channel of the microfluidic chip, and standing to form the agar gel capable of forming a chemical concentration gradient.
Example 4 preparation of microfluidic chip
1) Obtaining a micro-fluidic chip with a channel with a specific size, wherein the channel on the micro-fluidic chip comprises a concentration gradient forming part and sample injection parts arranged at two ends of the concentration gradient forming part, and the channel on the micro-fluidic chip has the depth of 90 mu m, the width of 1.5mm and the length of 1cm; preparing a PDMS layer with a concentration gradient forming part and a groove of a sample injection part by adopting a photoetching method, then punching a sample injection hole reaching a channel at the position of the sample injection part of the PDMS layer, wherein the diameter of the sample injection hole is 1.1mm, and then bonding with a glass slide;
2) Preparing an agar gel solution, wherein the concentration of the agar gel solution is 0.15%;
3) Injecting the agar gel solution into the channel of the microfluidic chip, standing to form the agar gel capable of forming chemical concentration gradient
As is clear from the state of the agar gels of comparative examples 1 to 4, the gels of examples 1 to 3 were uniform and stable, while the gel of example 4 was easily broken, indicating that the depth greatly affects the gel state and that it is necessary to control the depth of the channel to 80 μm or less.
Example 5 preparation of microfluidic chip
1) Obtaining a micro-fluidic chip with a channel with a specific size, wherein the channel on the micro-fluidic chip comprises a concentration gradient forming part and sample injection parts arranged at two ends of the concentration gradient forming part, and the channel on the micro-fluidic chip has the depth of 90 mu m, the width of 2.5mm and the length of 1cm; preparing a PDMS layer with a concentration gradient forming part and a groove of a sample injection part by adopting a photoetching method, and then punching a sample injection hole reaching a channel at the position of the sample injection part of the PDMS layer, wherein the diameter of the sample injection hole is 1.5mm and the sample injection hole is bonded with a glass slide;
2) Preparing an agar solution, wherein the concentration of the agar solution is 0.3%;
3) And injecting the agar solution into a channel of the microfluidic chip, and standing to form agar gel capable of forming chemical concentration gradient.
The gel state of example 5 was uniformly stable.
Example 6 application
The microfluidic chip prepared in example 1 was used, fluorescent dye was added to the sample wells, and the grey values were obtained by analyzing the color of the fluorescent dye, and further analyzed to evaluate whether a concentration gradient had formed.
The invention is used for establishing stable chemical concentration gradient, and the experimental result is the same as the expectation, and the gradient with different concentration distribution can be generated by adjusting the concentration of the agar. FIG. 2 shows a grey value analysis of the concentration gradient formed by the fluorescent dye in the channel. The chemical gradient is traced and displayed by the fluorescent dye, and the fluorescence intensity in the channel can be seen to be reduced from one end of the fluorescent dye injection to the other end in sequence under a microscope. The grey value is used to analyze the distribution data of the fluorescent dye to obtain the concentration change of the fluorescent dye in the channel, and fig. 3 shows that the fluorescent dye has a good concentration gradient. And the concentration gradient can be preserved for at least 12 hours. And can realize a concentration difference of 5% -100%.
Claims (10)
1. A method for preparing a microfluidic chip capable of setting chemical concentration gradient is characterized by comprising the following steps:
1) Obtaining a micro-fluidic chip with a channel with a specific size, wherein the channel on the micro-fluidic chip comprises a concentration gradient forming part and sample injection parts arranged at two ends of the concentration gradient forming part, and the depth of the channel on the micro-fluidic chip is less than 80 mu m; a channel on the microfluidic chip is punched to the channel through a PDMS layer with a concentration gradient forming part and a groove of a sample injection part, and a sample injection hole is formed in the PDMS layer at the position of the sample injection part; then bonding with a glass slide to form a channel containing a concentration gradient forming part and a sample injection part, and then
2) Preparing an agar gel solution, wherein the concentration of the agar solution is 0.15% -0.3%;
3) And injecting the agar solution into a channel of the microfluidic chip, and standing to form agar gel capable of forming chemical concentration gradient.
2. The preparation method according to claim 1, further comprising, in the step 4), injecting the active ingredient andor the dye through the sampling hole at one side of the channel, and obtaining the microfluidic chip with a concentration gradient after standing.
3. The method of claim 1, wherein the channels on the microfluidic chip have a rectangular cross-section.
4. The method of claim 1, wherein the depth of the channel on the microfluidic chip is 40 to 55 μm.
5. The method of claim 1, wherein the width of the channel on the microfluidic chip is 0.8 to 2.5mm, and the length of the channel on the microfluidic chip is 0.8 to 2.5cm.
6. The microfluidic chip capable of setting the chemical concentration gradient is characterized in that a channel is arranged on the microfluidic chip and comprises a concentration gradient forming part and sample injection parts arranged at two ends of the concentration gradient forming part, and the depth of the channel on the microfluidic chip is less than 80 microns; the channel on the microfluidic chip is bonded with the glass slide through the PDMS layer provided with the groove of the concentration gradient forming part and the sample injection part to form a sealed channel comprising the concentration gradient forming part and the sample injection part, and then the PDMS layer at the position of the sample injection part is punched to the channel to form a sample injection hole; and the channel also comprises agar gel, and the concentration of the agar gel is 0.15% -0.3%.
7. The microfluidic chip according to claim 6, wherein the number of the channels is more than one.
8. The microfluidic chip according to claim 6, characterized in that it is prepared by the preparation method according to any one of claims 1 to 5.
9. Use of a microfluidic chip according to any of claims 6 to 8 as a tracking device for microscopic microorganisms or cells.
10. A tracing device for microorganisms or cells, characterized in that it comprises a microfluidic chip according to any one of claims 6 to 8 and an observation or recording device, preferably selected from fluorescence inverted microscopes.
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| CN202110348997.XA CN115138402A (en) | 2021-03-31 | 2021-03-31 | Micro-fluidic chip capable of setting chemical concentration gradient and preparation method and application thereof |
| PCT/CN2021/138001 WO2022206023A1 (en) | 2021-03-31 | 2021-12-14 | Microfluidic chip capable of setting chemical concentration gradient, preparation method therefor and use thereof |
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