CN214142363U - Multi-channel micro-fluidic chip for analyzing cell migration characteristics - Google Patents
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Abstract
The utility model provides a multi-channel micro-fluidic chip for analyzing cell migration characteristics, which comprises a glass substrate and a PDMS chip main body closely attached to the glass substrate; a plurality of groups of cell migration and movement analysis units for generating different chemokine concentration gradient environments are arranged on the PDMS chip main body in parallel; each cell migration movement analysis unit includes: a cell loading unit for injecting cells and determining initial positions of the cells; the cell chemotaxis migration unit is used for adding cell culture solution and chemotactic factors and can form chemotactic factor concentration gradient to attract cells to generate migration movement; and a cell arresting unit for arresting the cells from crossing the initial cell location area when the concentration gradient is not formed. The utility model discloses after pouring into the neutrophil into, can simulate the directional migration phenomenon that takes place after the neutrophil in human capillary receives chemotactic factor stimulation, have certain detection flux.
Description
Technical Field
The utility model relates to a cell biology experimental apparatus especially relates to a cell migration characteristic is multichannel micro-fluidic chip for analysis.
Background
The micro-fluidic chip is a scientific technology which is mainly characterized by controlling fluid in a micron-scale space, and can integrate experimental operation units such as biochemistry, medicine and the like on a micron-scale chip. The microfluidic chip can realize flexible combination and scale integration of various unit operation technologies on an integrally controllable micro platform, becomes a cell research platform with the most development potential, and can be used for immune cell migration characteristic research. Neutrophils, a key cell of the innate immune system, play important roles in the body's immune response against infection through chemotaxis and phagocytosis. This is because when the body is infected or inflamed, neutrophils activate polarization for the first time and extravasate the vessel wall, then migrate towards chemokines released from the foci of infection, and finally recognize and phagocytose pathogenic bacteria, while releasing Reactive Oxygen Species (ROS) and proteolytic enzymes to kill bacteria and microorganisms. Therefore, the research on the chemotactic movement of the neutrophils and the whole process of the neutrophil phagocytosis has very important scientific significance for monitoring the immune function state of the chronic patients such as diabetes and the like and evaluating the curative effect.
Common methods for evaluating the cell migration characteristics mainly comprise a scratch experiment method and a transwell experiment method, but the two methods have obvious defects, mainly show that the scratch experiment method has poor repeatability and damages cells. The Transwell's rule of experiment is complex to operate and is not suitable for dynamic observation. For this reason, microfluidic gradient generators based on microfluidic chips have been developed. The size of the pipeline of the microfluidic chip is in the micron order, the size of the pipeline is equivalent to that of a cell, the consumed cell and reagent amount is small, the in-vivo environment can be well simulated, the cell is not damaged, and the microfluidic chip is suitable for dynamic observation. However, the current research mostly uses single-channel microfluidic chips, and has the defects of low flux, poor integration level, poor repeatability among chips and the like. In addition, cell migration characteristic research based on a microfluidic chip usually depends on professional living cell imaging equipment, and after a series of cell images are collected at regular time, professional researchers need to manually track cell movement tracks and analyze cell chemotaxis by using ImageJ image processing software, so that the detection method is complex, and the time and experiment cost are high. In addition, the subjective awareness of different experimenters can seriously affect the accuracy and repeatability of the data of the analysis of the chemotactic migration of cells.
SUMMERY OF THE UTILITY MODEL
The utility model provides a cell migration is multichannel micro-fluidic chip for characteristic analysis to solve the problem that present single channel micro-fluidic chip flux is low, the integrated level is poor, repeatability is poor between the chip, with after pouring into the neutrophil into, can simulate the directional migration phenomenon that takes place after the neutrophil in the human capillary receives chemotactic factor stimulation, have certain detection flux.
The technical scheme of the utility model is realized like this:
a multi-channel micro-fluidic chip for analyzing cell migration characteristics comprises a glass substrate and a PDMS chip main body tightly attached to the glass substrate; a plurality of groups of cell migration and movement analysis units for generating different chemokine concentration gradient environments are arranged on the PDMS chip main body in parallel; each cell migration movement analysis unit includes:
a cell loading unit for injecting cells and determining initial positions of the cells;
the cell chemotaxis migration unit is used for adding cell culture solution and chemotactic factors and can form chemotactic factor concentration gradient to attract cells to generate migration movement; and
and the cell blocking unit is used for blocking the cells from crossing the initial position locating area of the cells when the concentration gradient is not formed.
According to a further optimized technical scheme, the cell loading unit comprises:
a cell loading port for injecting cells;
the cell loading pipeline is communicated with the cell loading port; and
a cell initial position locating area, which can be reached by the cell along the cell loading pipeline.
In a further preferred embodiment, the cell chemotaxis migration unit comprises:
a cell culture solution loading port for adding a cell culture solution;
a chemokine loading port for adding a chemokine;
the snakelike pipeline is connected with the cell culture solution loading port through the cell migration movement pipeline and is connected with the chemotactic factor loading port through the cell culture solution pipeline, and the added chemotactic factor and the cell culture solution can balance the pressure difference between the two sides through the snakelike pipeline;
the chemotactic factor and the cell culture solution after the pressure difference between the two parts is balanced by the snake-shaped pipeline can be converged into the cell migration movement pipeline to form a stable concentration gradient in the cell migration movement pipeline; and
and the waste liquid outlet is connected with the cell migration movement pipeline and can discharge waste liquid.
According to the technical scheme, four cell migration and movement analysis units are arranged in parallel.
A method of analyzing cell migration characteristics, comprising the steps of:
s1, preparing a microfluidic chip, separating neutrophils, and preparing glucose solution, chemokine solution, cell culture solution and fibrinectin solution with different concentrations;
s2, placing the neutrophils into glucose solutions with different concentrations for cultivation;
s3, paving the micro-fluidic chip pipeline in a fibrinectin solution for placement;
s4, sucking out the fibrinectin solution, and paving the micro-fluidic chip pipeline with the cell culture solution;
s5, placing the microfluidic chip into a microscope system, adjusting the focal length, finding a cell migration movement area, and setting the image acquisition interval time and number;
s6, injecting the separated neutrophils into the cell injection port by using a pipette gun;
s7, after the neutrophils are arranged in the initial cell position positioning area, a liquid transfer gun is used for taking the chemotactic factors and injecting the chemotactic factors into a chemotactic factor injection opening; meanwhile, a pipette is used for taking the cell culture solution and injecting the cell culture solution into a cell culture solution injection port;
s8, flowing the reagents in the chemotactic factor injection port and the cell culture solution injection port along the chemotactic factor injection pipeline and the cell culture solution pipeline, and after the stable pressure difference in the serpentine pipeline, flowing the reagents in the cell migration movement pipeline, and constructing a stable concentration gradient environment;
s9, after the cells sense the existence of the chemotactic factors, the cells penetrate through the cell blocking unit and enter a cell migration movement pipeline from the initial position positioning area of the cells;
the chemotaxis migration movement state of the cells in the environment of different chemokine concentration gradients is recorded and analyzed by the chemotaxis migration analysis system at the same time.
Further optimizing the technical scheme, when cell migration characteristic analysis is carried out, the cell chemotaxis is simply analyzed by dividing the width pixels of the first cell image and the last cell image into 10 parts and adopting a cell partition counting and digital scoring method.
And further optimizing the technical scheme, and when cell migration characteristic analysis is carried out, tracking the cell movement trace by identifying the center coordinates of the cells and combining a minimum distance method, thereby calculating the chemotaxis index, the total migration distance, the movement speed and the gradient displacement of the cells.
By adopting the technical scheme, the beneficial effects of the utility model are that:
the utility model discloses can observe simultaneously under the different concentration gradient the chemotaxis migration motion condition of neutrophil in micro-fluidic chip, have certain experiment flux. The utility model discloses after pouring into the neutrophil into, can simulate the directional migration phenomenon that takes place after the neutrophil in the human capillary receives chemotactic factor stimulation, have certain detection flux, solved present single channel micro-fluidic chip flux low, integrated level poor, the poor problem of repeatability between the chip effectively.
The utility model discloses but the automatic analysis cell migration state, this micro-fluidic chip and system can be used to all kinds of cell chemotaxis migration research. The utility model discloses liquid in chemokine loading mouth and cell culture liquid loading mouth can form stable concentration gradient in cell migration motion region. After the cell senses the existence of the chemotactic factor, the cell passes through the cell blocking area and then enters the cell migration movement area from the initial position positioning area of the cell. The cell migration movement region can be placed under a microscope observation device to observe the migration movement characteristics of the cells. The chemotaxis migration movement state of the cells in the environment of 4 different chemokine concentration gradients can be recorded and analyzed simultaneously.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without inventive exercise.
Fig. 1 is a schematic structural diagram of a multi-channel microfluidic chip for analyzing cell migration characteristics according to the present invention;
fig. 2 is a schematic structural diagram of a cell migration movement analysis unit i in a multi-channel microfluidic chip for analyzing cell migration characteristics according to the present invention;
FIG. 3 is a schematic structural diagram of a second cell migration movement analysis unit in the multi-channel microfluidic chip for cell migration characteristic analysis according to the present invention;
fig. 4 is a schematic structural diagram of a cell migration movement analysis unit iii in the multi-channel microfluidic chip for analyzing cell migration characteristics according to the present invention;
fig. 5 is a schematic structural diagram of a cell migration movement analysis unit four in the multi-channel microfluidic chip for analyzing cell migration characteristics according to the present invention;
FIG. 6 is a schematic diagram of the structure of four cell migration and movement analysis units of the four-channel chip of the present invention;
FIG. 7 is a diagram of the conclusion of hyperglycemia-identical FMLPs of the present invention;
FIG. 8 is a graph of the results of different FMLPs from normal cells of the present invention;
FIG. 9 shows the migration of neutrophils at 0min and 15min after glucose treatment at 0mM/L, 1mM/L, 5mM/L and 25mM/L, respectively, from left to right.
Wherein:
11. the cell migration and movement analysis unit I, 11(1), the cell loading unit I, 11(1) a, the cell injection port I, 11(1) b, the cell injection pipeline I, 11(2), the cell chemotaxis unit I, 11(2) a, the chemotactic factor injection port I, 11(2) b, the cell culture solution injection port I, 11(2) c, the chemotactic factor injection pipeline I, 11(2) d, the cell culture solution injection pipeline I, 11(2) e, the snake-shaped pipeline I, 11(2) f, the cell migration and movement pipeline I, 11(2) g, the waste liquid discharge port I, 11(3) and the cell blocking unit I;
12. a second cell migration and movement analysis unit, a second cell loading unit, a 12(1) a, a second cell injection port, a 12(1) b, a second cell injection pipeline, a 12(2), a second cell chemotaxis unit, a 12(2) a, a second chemotactic factor injection port, a second cell culture solution injection port, a 12(2) c, a second chemotactic factor injection pipeline, a 12(2) d, a second cell culture solution injection pipeline, a 12(2) e, a second snake-shaped pipeline, a 12(2) f, a second cell migration and movement pipeline, a 12(2) g, a second waste liquid discharge port, a 12(3) and a second cell blocking unit;
13. a cell migration movement analysis unit III, 13(1), a cell loading unit III, 13(1) a, a cell injection port III, 13(1) b, a cell injection pipeline III, 13(2), a cell chemotaxis unit III, 13(2) a, a chemotactic factor injection port III, 13(2) b, a cell culture solution injection port III, 13(2) c, a chemotactic factor injection pipeline III, 13(2) d, a cell culture solution injection pipeline III, 13(2) e, a snake-shaped pipeline III, 13(2) f, a cell migration movement pipeline III, 13(2) g, a waste liquid discharge port III, 13(3) and a cell blocking unit III;
14. the cell migration movement analysis unit four, 14(1), the cell loading unit four, 14(1) a, the cell injection port four, 14(1) b, the cell injection pipeline four, 14(2), the cell chemotaxis unit four, 14(2) a, the chemotactic factor injection port four, 14(2) b, the cell culture fluid injection port four, 14(2) c, the chemotactic factor injection pipeline four, 14(2) d, the cell culture fluid injection pipeline four, 14(2) e, the serpentine pipeline four, 14(2) f, the cell migration movement pipeline four, 14(2) g, the waste fluid discharge port four, 14(3) and the cell blocking unit four.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.
Example 1
A multi-channel micro-fluidic chip for analyzing cell migration characteristics is shown in a combined manner in figures 1 to 6 and comprises a glass substrate and a PDMS chip main body tightly attached to the glass substrate, wherein the PDMS chip main body and the glass substrate are tightly attached to each other after being processed by a plasma bonding process.
The PDMS chip main body is provided with a plurality of groups of cell migration and movement analysis units which are used for generating different chemokine concentration gradient environments in parallel. The utility model discloses well cell migration motion analysis unit is provided with four side by side, can set up four different chemotactic factor concentration gradient environments of group simultaneously.
Each cell migration movement analysis unit includes: cell loading unit, cell chemotaxis migration unit, cell arresting unit.
And the cell loading unit is used for injecting cells and determining initial positions of the cells.
And the cell chemotaxis migration unit is used for adding cell culture solution and chemotactic factors and can form a chemotactic factor concentration gradient to attract cells to perform migration movement.
And the cell blocking unit is used for blocking the cells from crossing the initial position locating area of the cells when the concentration gradient is not formed.
The cell loading unit comprises: a cell loading port, a cell loading pipeline and a cell initial position positioning area.
A cell loading port for injecting cells.
And the cell loading pipeline is communicated with the cell loading port.
A cell initial position locating area, which can be reached by the cell along the cell loading pipeline.
The distance between the cell blocking unit and the glass substrate is 5 mu m, which is slightly smaller than the diameter of the cell, and the injected cell is blocked in the cell initial position positioning area by the cell blocking unit.
The cell chemotaxis migration unit comprises: cell culture liquid loading mouth, chemotactic factor loading mouth, cell culture liquid pipeline, chemotactic factor pipeline, waste liquid export, cell migration motion pipeline.
A cell culture solution loading port for adding a cell culture solution;
a chemokine loading port for adding a chemokine;
the snakelike pipeline is connected with the cell culture solution loading port through the cell migration movement pipeline and is connected with the chemotactic factor loading port through the cell culture solution pipeline, and the added chemotactic factor and the cell culture solution can balance the pressure difference between the two sides through the snakelike pipeline;
the chemotactic factor and the cell culture solution after the pressure difference between the two parts is balanced by the snake-shaped pipeline can be converged into the cell migration movement pipeline to form a stable concentration gradient in the cell migration movement pipeline; and the waste liquid outlet is connected with the cell migration movement pipeline and can discharge waste liquid.
The liquid in the chemotactic factor loading port and the cell culture solution loading port flows in along the chemotactic factor pipeline and the cell culture solution pipeline, and is converged into the cell migration movement pipeline after the pressure difference between the chemotactic factor pipeline and the cell culture solution pipeline is balanced through the snake-shaped pipeline, so that a stable concentration gradient is formed in the cell migration movement pipeline.
After the cell senses the existence of the chemotactic factor, the cell passes through the cell blocking unit and enters a cell migration movement pipeline from the initial position positioning area of the cell. The cell migration movement pipeline can be arranged below a microscope observation device to observe the migration movement characteristics of the cells. The chemotaxis migration movement state of the cells in the environment of 4 different chemokine concentration gradients can be recorded and analyzed simultaneously.
Example 2
The embodiment discloses a cell migration characteristic analysis method, which comprises the following steps:
s1, preparing a microfluidic chip, separating neutrophils, and preparing glucose solution, chemokine solution, cell culture solution and fibrinectin solution with different concentrations.
S2, placing the neutrophils into glucose solutions with different concentrations for incubation for one hour.
And S3, paving the pipeline of the microfluidic chip in a fibrinectin solution for standing for forty-five minutes.
S4, sucking out the fibrinectin solution, paving the micro-fluidic chip pipeline with the cell culture solution, and standing for forty-five minutes.
And S5, placing the microfluidic chip into a microscope system, adjusting the focal length, finding a cell migration movement area, and setting the image acquisition interval time and number.
S6, the separated neutrophils are injected into four cell injection ports, namely, a first cell injection port 11(1) a, a second cell injection port 12(1) a, a third cell injection port 13(1) a and a fourth cell injection port 14(1) a by using a pipette.
S7, after the neutrophils are arranged in the initial cell location area, 100 μ L of the chemokine is taken by a pipette and injected into four chemokine injection ports, namely a chemokine injection port I11 (2) a, a chemokine injection port II 12(2) a, a chemokine injection port III 13(2) a and a chemokine injection port IV 14(4) a. Meanwhile, 100. mu.L of cell culture fluid is taken by a pipette and injected into four cell culture fluid injection ports, namely, a first cell culture fluid injection port 11(2) b, a second cell culture fluid injection port 12(2) b, a third cell culture fluid injection port 13(2) b and a fourth cell culture fluid injection port 14(2) b.
S8, the reagents in the chemokine injection port and the cell culture fluid injection port flow in along the chemokine injection pipeline and the cell culture fluid pipeline, and flow into the cell migration movement pipeline one 11(2) f, the cell migration movement pipeline two 12(2) f, the cell migration movement pipeline three 13(2) f and the cell migration movement pipeline four 14(2) f after stable pressure difference in the serpentine pipeline one 11(2) e, the serpentine pipeline two 12(2) e, the serpentine pipeline three 13(2) e and the serpentine pipeline four 14(2) e, and a stable concentration gradient environment is constructed.
S9, the cell will pass through the cell blocking unit after sensing the existence of the chemotactic factor, and enter the cell migration moving pipe from the initial position positioning area of the cell.
The chemotaxis migration movement state of the cells in the environment of different chemokine concentration gradients is recorded and analyzed by the chemotaxis migration analysis system at the same time.
In the analysis of cell migration characteristics, cell chemotaxis was simply analyzed by cell division counting and numerical scoring by dividing the width pixels of the first and last cell images into 10 parts.
When the cell migration characteristic analysis is carried out, the chemotaxis index, the total migration distance, the movement speed and the gradient displacement of the cell can be further calculated by identifying the central coordinates of the cell and tracking the movement trace of the cell by combining a minimum distance method.
Example 3
This example discloses the chemotactic effect of hyperglycemia on neutrophils, comprising the following steps:
step 1: a microfluidic chip was prepared, neutrophils were separated, and 0mM/L, 1mM/L, 5mM/L, and 25mM/L of glucose, 0.4% BSA (bovine serum albumin) solution, 1g/L of fibrinectin solution, and 10nM/L of fMLP (N-formylmethionyl-leucyl-phenylalanine) solution were prepared.
Step 2: and (3) paving the pipeline of the microfluidic chip with the fibrinectin solution, and standing for forty-five minutes.
And step 3: the fibrinectin solution was aspirated, the microfluidic chip tubing was filled with 0.4% BSA solution, and left for forty-five minutes.
And 4, step 4: put into conventional microscope system with the micro-fluidic chip perhaps the utility model provides a portable cell chemotaxis migration analytic system goes up and adjusts the focus, finds the cell migration motion region. And simultaneously setting the time and the number of image acquisition intervals.
And 5: the separated neutrophils are injected into four cell injection ports, namely a cell injection port I1 (1) a, a cell injection port II 2(1) a, a cell injection port III 3(1) a and a cell injection port IV 4(1) a, by a pipette.
Step 6: after the neutrophils were aligned on the primary localization area of the cells, 100. mu.L of 10nM/LfMLP was injected into the chemokine injection ports one 11(2) a, two 12(2) a, three 13(2) a, and four 14(4) a, respectively, using a pipette gun. At the same time, 100. mu.L of 0.4% BSA solution was injected into the first cell culture fluid inlet 11(2) b, the second cell culture fluid inlet 12(2) b, the third cell culture fluid inlet 13(2) b, and the fourth cell culture fluid inlet 14(4) b by using a pipette.
And 7: the fMLP solution and the BSA solution are flowed along the chemokine injection line and the cell culture liquid line, and after a stable pressure difference is maintained in the first serpentine line (11) (2) e, the second serpentine line (12) (2) e, the third serpentine line (13) (2) e, and the fourth serpentine line (14) (2) e, the fMLP solution and the BSA solution are flowed into the first cell migration line (11) (2) f, the second cell migration line (12) (2) f, the third cell migration line (13) (2) f, and the fourth cell migration line (14) (2) f, and a stable concentration gradient environment is constructed.
And 8: and installing the microfluidic chip on a portable cell chemotaxis migration analysis system, and starting a camera to acquire a real-time moving image of the cells in the microfluidic chip and performing cell chemotaxis migration behavior analysis.
And step 9: the chemotaxis speed V of the neutrophil is processed by image acquisition, image preprocessing, a Canny algorithm, a cell image partition technical algorithm and a cell image partition technical algorithm by cell chemotaxis migration analysis system softwareMSChemotaxis index CI, gradient direction displacement LGDTotal migration distance LADCalculations were performed to assess cell chemotactic activity.
Example 4
This example discloses the effect of various concentrations of chemokine (fMLP) on neutrophil chemotaxis, comprising the steps of:
step 1: preparing a microfluidic chip, separating neutrophils, and preparing 0nM/L, 1nM/L, 10nM/L, 100nM/LfMLP, 0.4% BSA solution and 1g/L fibrinectin solution.
Step 2: and (3) paving the pipeline of the microfluidic chip with the fibrinectin solution, and standing for forty-five minutes.
And step 3: the fibrinectin solution was aspirated, the microfluidic chip tubing was filled with 0.4% BSA solution, and left for forty-five minutes.
And 4, step 4: put into conventional microscope system with the micro-fluidic chip perhaps the utility model provides a portable cell chemotaxis migration analytic system goes up and adjusts the focus, finds the cell migration motion region. And simultaneously setting the time and the number of image acquisition intervals.
And 5: the separated neutrophils are injected into four cell injection ports, namely a cell injection port I1 (1) a, a cell injection port II 2(1) a, a cell injection port III 3(1) a and a cell injection port IV 4(1) a, by a pipette.
Step 6: after the neutrophils were aligned on the primary localization area of the cells, 100. mu.L of 0nM/L, 1nM/L, 10nM/L, 100nM/L fMLP solution was injected into the chemokine injection port one 11(2) a, the chemokine injection port two 12(2) a, the chemokine injection port three 13(2) a, the chemokine injection port four 14(4) a four chemokine injection ports, respectively, using a pipette gun. At the same time, 100. mu.L of 0.4% bovine serum albumin was injected into the first cell culture fluid inlet 11(2) b, the second cell culture fluid inlet 12(2) b, the third cell culture fluid inlet 13(2) b, and the fourth cell culture fluid inlet 14(4) b by using a pipette.
And 7: the reagents in the chemokine injection port and the cell culture fluid injection port flow in along the chemokine injection pipeline and the cell culture fluid pipeline, and flow into the cell migration movement pipeline one 11(2) f, the cell migration movement pipeline two 12(2) f, the cell migration movement pipeline three 13(2) f and the cell migration movement pipeline four 14(2) f after stable pressure difference in the serpentine pipeline one 11(2) e, the serpentine pipeline two 12(2) e, the serpentine pipeline three 13(2) e and the serpentine pipeline four 14(2) e, and a stable concentration gradient environment is constructed.
And 8: and installing the microfluidic chip on a portable cell chemotaxis migration analysis system, and starting a camera to acquire a real-time moving image of the cells in the microfluidic chip and performing cell chemotaxis migration behavior analysis.
And step 9: the chemotaxis speed V of the neutrophil is processed by image acquisition, image preprocessing, a Canny algorithm, a cell image partition technical algorithm and a cell image partition technical algorithm by cell chemotaxis migration analysis system softwareMSChemotaxis index CI, gradient direction displacement LGDTotal migration distance LADCalculations were performed to assess cell chemotactic activity.
The above description is only a preferred embodiment of the present invention, and should not be taken as limiting the invention, and any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (4)
1. A multi-channel micro-fluidic chip for analyzing cell migration characteristics is characterized by comprising a glass substrate and a PDMS chip main body which is tightly attached to the glass substrate; a plurality of groups of cell migration and movement analysis units for generating different chemokine concentration gradient environments are arranged on the PDMS chip main body in parallel; each cell migration movement analysis unit includes:
a cell loading unit for injecting cells and determining initial positions of the cells;
a cell chemotactic migration unit for adding a cell culture solution and a chemotactic factor and capable of forming a concentration gradient of the chemotactic factor to attract cells to undergo migration movement, and
a cell-arresting unit for arresting the cells across the initially localized area of the cells when the concentration gradient is not formed.
2. The multi-channel microfluidic chip for cell migration characteristic analysis according to claim 1, wherein the cell loading unit comprises:
a cell loading port for injecting cells;
a cell loading conduit in communication with the cell loading port, an
A cell initial position locating area, which can be reached by the cell along the cell loading pipeline.
3. The multi-channel microfluidic chip for cell migration characteristic analysis according to claim 1, wherein said cell chemotaxis migration unit comprises:
a cell culture fluid loading port for adding a cell culture fluid;
a chemokine loading port for adding a chemokine;
the snakelike pipeline is connected with the cell culture solution loading port through the cell migration movement pipeline and is connected with the chemotactic factor loading port through the cell culture solution pipeline, and the added chemotactic factor and the cell culture solution can balance the pressure difference between the two sides through the snakelike pipeline;
the chemotactic factor and the cell culture solution after the pressure difference between the two parts is balanced by the snake-shaped pipeline can be converged into the cell migration movement pipeline to form a stable concentration gradient in the cell migration movement pipeline; and
and the waste liquid outlet is connected with the cell migration movement pipeline and can discharge waste liquid.
4. The multi-channel microfluidic chip for cell migration characteristic analysis according to claim 1, wherein four cell migration movement analysis units are arranged in parallel.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112501005A (en) * | 2020-12-21 | 2021-03-16 | 合肥中科易康达生物医学有限公司 | Multi-channel micro-fluidic chip and method for analyzing cell migration characteristics |
| CN113980794A (en) * | 2021-09-22 | 2022-01-28 | 中国科学院合肥物质科学研究院 | Multi-channel micro-fluidic chip suitable for cell migration analysis and application thereof |
| CN114917754A (en) * | 2022-05-06 | 2022-08-19 | 东南大学 | Microfluidic colloidal particle separation device and separation method |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112501005A (en) * | 2020-12-21 | 2021-03-16 | 合肥中科易康达生物医学有限公司 | Multi-channel micro-fluidic chip and method for analyzing cell migration characteristics |
| CN113980794A (en) * | 2021-09-22 | 2022-01-28 | 中国科学院合肥物质科学研究院 | Multi-channel micro-fluidic chip suitable for cell migration analysis and application thereof |
| CN113980794B (en) * | 2021-09-22 | 2024-04-16 | 中国科学院合肥物质科学研究院 | Multichannel microfluidic chip suitable for cell migration analysis and application thereof |
| CN114917754A (en) * | 2022-05-06 | 2022-08-19 | 东南大学 | Microfluidic colloidal particle separation device and separation method |
| CN114917754B (en) * | 2022-05-06 | 2023-02-17 | 东南大学 | Microfluidic colloidal particle separation device and separation method |
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