CN217499256U

CN217499256U - Single cell capture micro-fluidic chip

Info

Publication number: CN217499256U
Application number: CN202123352288.3U
Authority: CN
Inventors: 李长明; 史转转
Original assignee: Suzhou University of Science and Technology
Current assignee: Suzhou University of Science and Technology
Priority date: 2021-12-28
Filing date: 2021-12-28
Publication date: 2022-09-27
Anticipated expiration: 2031-12-28

Abstract

The utility model discloses a micro-fluidic chip is caught to unicell, micro-fluidic chip includes a plurality of concave type microfluid passageway, the highest point bottom of microfluid passageway is provided with the micropore, and set up the detection cell below the micropore, the microfluid passageway just is provided with the microfluid branch road perpendicular to microfluid passageway above the micropore, the detection cell bottom communicates with the lower of microfluid passageway, detect the electrode of cell real-time supervision of bottom embedding of cell, cell suspension flows through the chip main entrance with certain velocity of flow during the detection, realize high flux unicell through cross-like microfluid's control and micropore's form and catch, other external forces need not be exerted in the capture process; meanwhile, the electrode positioned at the bottom of the detection pool carries out real-time sensing analysis on the state of the cell and the released small biological molecules, which has important significance on real-time and rapid capture and analysis of the single cell and provides a high-efficiency and stable multifunctional analysis platform for the fields of single cell sequencing, single cell real-time analysis and the like.

Description

Single cell capture micro-fluidic chip

Technical Field

The utility model relates to a unicellular detection field, concretely relates to micro-fluidic chip is caught to unicellular.

Background

Capturing single cells is an important prerequisite for the development of accurate tests at the single cell level. Currently, the optical tweezers trapping technology has certain advantages in the aspect of controlling single cells, and the optical tweezers have the capacity of positioning in a micron range and can accurately trap and move the single cells. The optical tweezers do not need to contact cells, the whole operation can be carried out in a sealed container, and the damage to the cells is less. However, optical tweezers require high equipment requirements, are expensive, and do not enable high throughput single cell capture and analysis. The micro-fluidic chip has the advantages of small volume, small sample consumption, high operation speed, high flux, low cost and the like, and can effectively avoid the error of manual operation so that the obtained data is more reliable. In addition, the processing material of the microfluidic chip has good biocompatibility and high transparency, and is suitable for observing the moving and capturing conditions of single cells. Therefore, the manipulation of single cells can be realized by designing a micron-sized two-dimensional or three-dimensional channel matched with the size of the cells in the microfluidic chip, and a new idea can be provided for meeting the requirement of the single cell analysis field on the single cell capture by using the microfluidic chip for capturing the single cells.

The traditional optical tweezers trapping technology can realize the control of single cells and can accurately trap and move the single cells. However, the optical tweezers technology has high requirements on equipment and high price, and more importantly, high-throughput single cell capture and single cell analysis cannot be realized. In order to realize efficient and accurate high-throughput single-cell capture, most of the traditional capture methods are based on optical, electric, acoustic and magnetic methods, and the cells need to be driven to a specific capture position by means of external force. In many of the microfluidic single cell capture technologies reported at present, a platform based on dielectrophoresis and other technologies combined with microfluidics can realize high-throughput single cell capture, but the platform needs to regulate and control various operation parameters and cannot realize real-time detection and analysis of captured single cells. The single cell capture method based on droplet microfluidics is suitable for analysis of single cells by independently encapsulating single cells and other reaction reagents in micro-droplets, but the method often has the condition that no cell or a plurality of cells are encapsulated in a single droplet, and the position of the micro-droplet after encapsulating the single cells is difficult to fix and is not suitable for real-time observation of specific single cells. Therefore, a platform for efficiently observing single cells in real time is urgently needed.

SUMMERY OF THE UTILITY MODEL

In view of this, an object of the present invention is to provide a single-cell capture microfluidic chip, which reasonably designs the micro-structure in the chip, so that when a cell suspension flows through a main channel of the chip at a certain flow rate, the cell suspension can be captured by high-flux single-cell through the control of cross-like microfluid and the form of micro-pores, and no other external force is applied during the capture process. Meanwhile, the bottom of the detection pool below the micropores is provided with a microelectrode which can carry out real-time sensing analysis on the state of the cells and the released small biological molecules, so that the microelectrode has important significance on real-time and rapid capture and analysis of single cells, and a high-efficiency and stable multifunctional analysis platform is provided for the fields of single cell sequencing, single cell real-time analysis and the like.

In order to achieve the purpose, the invention provides the following technical scheme:

1. the single cell capturing micro-fluidic chip comprises a plurality of concave micro-fluidic channels, wherein the bottoms of the highest positions of the micro-fluidic channels are provided with micropores, a detection pool is arranged below the micropores, micro-fluidic branches perpendicular to the micro-fluidic channels are arranged at positions, right facing the micropores, of the micro-fluidic channels, the bottom of the detection pool is communicated with the lowest position of the micro-fluidic channels, electrodes for monitoring cells in real time are embedded into the bottom of the detection pool, and the diameters of the micro-fluidic channels or the micropores are larger than or equal to the diameter of a single cell and smaller than the diameters of two cells.

Preferably, the diameter of the microporous microfluidic branch is greater than or equal to the diameter of a single cell and less than the diameter of two cells.

Preferably, the diameter of the channel connecting the detection cell and the microfluidic channel is smaller than the diameter of a single cell.

Preferably, the electrode is selected from, but not limited to, a gold electrode, a carbon electrode, an ITO electrode.

Preferably, the width range of the microfluidic channel is 15-45 μm; the diameter of the micropores is 25-60 mu m.

Preferably, the diameter of a channel connecting the detection cell and the microfluidic channel is 5-25 μm.

According to the invention, the diameter of the microfluidic branch is preferably 10-30 μm.

When the micro-fluidic chip is used for detection, cell suspension flows into the micro-fluidic channel at a certain speed, when the cell suspension passes through the upper part of the micro-hole along the micro-fluidic channel, a buffer solution is introduced into the micro-fluidic branch to extrude a single cell into the micro-hole under the action of micro-fluid, and then the electrode in the detection cell monitors the cell in real time.

The beneficial effects of the utility model reside in that: the utility model discloses a single cell capture micro-fluidic chip, which realizes high flux single cell capture through the control of cross-like microfluid and micropore mode when cell suspension flows through a chip main channel at a certain flow speed by reasonably designing the micro structure in the chip, and other external force is not required to be applied in the capture process; meanwhile, the bottom of the detection pool below the micropores is provided with a microelectrode which can carry out real-time sensing analysis on the state of the cells and the released small biological molecules, so that the microelectrode has important significance on real-time and rapid capture and analysis of single cells, and a high-efficiency and stable multifunctional analysis platform is provided for the fields of single cell sequencing, single cell real-time analysis and the like.

Drawings

In order to make the purpose, technical scheme and beneficial effect of the utility model clearer, the utility model provides a following figure explains:

FIG. 1 is a diagram of a microfluidic chip for high throughput single cell capture and real-time analysis;

fig. 2 is a schematic diagram of the structural dimensions of the microchannel (D denotes the width of the microfluidic channel; W denotes the vertical channel above the microwell; D is the diameter of the microwell; D denotes the width of the lower outlet; L ═ 30 μm denotes the distance from the center of the microwell to the center line of the microfluidic).

Detailed Description

The present invention will be further described with reference to the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and can implement the present invention, but the embodiments are not to be construed as limiting the present invention.

Example 1 Single cell-trapping microfluidic chip

The structure of the single-cell-capturing microfluidic chip is shown in figure 1. The micro-fluidic chip comprises a plurality of concave micro-fluidic channels 1, micropores 2 capable of passing single cells are arranged at the bottoms of the highest positions of the concave micro-fluidic channels, detection pools 3 are arranged below the micropores, micro-fluidic branches 4 perpendicular to the micro-fluidic channels are arranged at positions, right opposite to the micropores 2, of the micro-fluidic channels, the bottoms of the detection pools are communicated with the lowest positions of the micro-fluidic channels, electrodes 5 for monitoring cells in real time are embedded into the bottoms of the detection pools, and the diameters of the micro-fluidic channels or the micropores are larger than or equal to the diameter of a single cell and smaller than the diameters of two cells. When the microfluidic chip is used for detection, cell suspension flows into the microfluidic channel at a certain speed, when the cell suspension passes through the upper part of the micropores along the microfluidic channel, a buffer solution is introduced into the microfluidic branch to extrude a single cell into the micropore under the action of the microfluidic channel, and then the electrode in the detection cell monitors the cell in real time, and the monitoring is finished. After the detection is completed, the cells are washed out by back-flushing.

The micro-fluidic chip can be designed with a micro-channel structure with the same size as the cell, can control micro-fluid, is suitable for high-flux single cell capture, and can integrate various operation units for single cell capture and single cell real-time monitoring, thereby providing an efficient and low-cost analysis platform for single cell analysis. Specifically, the width range of the microfluidic channel is 15-45 μm, the size of the microfluidic channel is adjusted according to the size of the cell, so that the width of the channel is larger than the diameter of a single cell and smaller than the diameters of two cells, the diameter range of the micropore is 25-60 μm, the width of an upper outlet of the micropore is larger than or equal to the diameter of the single cell and smaller than the diameters of two cells, the width of a lower outlet of the micropore is 5-25 μm (smaller than the diameter of the single cell), the width range of a vertical channel above the micropore is 10-30 μm, and when the cell passes through the upper part of the micropore, the cell is extruded into the micropore by controlling the microfluidic speed in the vertical direction (figure 2).

More preferably, the width of the microfluidic channel is in the range of 30 μm, the diameter of the microwell is in the range of 45 μm, wherein the width of the upper outlet of the microwell is equal to or greater than the diameter of a single cell and less than two cells, the width of the lower outlet is 10 μm (less than the diameter of a single cell), the width of the vertical channel above the microwell is in the range of 15 μm, and the cells are "squeezed" into the microwell by controlling the microfluidic speed in the vertical direction when passing over the microwell.

The designed single cell capture microfluidic chip can be designed to meet the micropores of different target capture cell numbers, and according to different monitoring requirements, the microfluidic chip system designs the electrodes at the bottoms of the micropores into sensing devices with different detection functions. The microfluidic chip comprises a plurality of operation units required by single cell analysis, and can simultaneously design and integrate single cell high-throughput capture and detection analysis units and the like on a tiny chip platform. The single cell capturing process on the microfluidic chip can be observed in real time through a microscope, and the requirements of simple and efficient operation and cell activity guarantee are met. The whole process is easy to control, and the application range is wider.

Furthermore, a microelectrode can be designed at the bottom of the micropore, a technical platform can be provided for a plurality of life science fields such as analysis of cell expression information and biomolecule release of cells, and the shape and the size of the microelectrode can be designed according to different application fields.

The above-mentioned embodiments are merely preferred embodiments for fully illustrating the present invention, and the scope of the present invention is not limited thereto. Equivalent substitutes or changes made by the technical personnel in the technical field on the basis of the utility model are all within the protection scope of the utility model. The protection scope of the present invention is subject to the claims.

Claims

1. The single cell capture micro-fluidic chip is characterized in that: the microfluidic chip comprises a plurality of concave microfluidic channels, micropores are arranged at the bottoms of the highest positions of the microfluidic channels, detection pools are arranged below the micropores, microfluidic branches perpendicular to the microfluidic channels are arranged above the positions, right opposite to the micropores, of the microfluidic channels, the bottoms of the detection pools are communicated with the lowest positions of the microfluidic channels, electrodes for monitoring cells in real time are embedded into the bottoms of the detection pools, and the diameters of the microfluidic channels or the micropores are larger than or equal to the diameter of a single cell and smaller than the diameters of two cells.

2. The single-cell-capture microfluidic chip of claim 1, wherein: the diameter of the microporous microfluidic branch is greater than or equal to the diameter of a single cell and less than the diameter of two cells.

3. The single-cell-capture microfluidic chip of claim 1, wherein: the diameter of the channel connecting the detection cell and the microfluidic channel is smaller than that of a single cell.

4. The single-cell-capture microfluidic chip of claim 1, wherein: the electrode is selected from one of a gold electrode, a carbon electrode and an ITO electrode.

5. The single-cell-capture microfluidic chip of claim 1, wherein: the width range of the microfluidic channel is 15-45 μm; the diameter of the micropores is 25-60 mu m.

6. The single-cell-capture microfluidic chip of claim 1, wherein: the diameter of a channel connecting the detection cell and the microfluidic channel is 5-25 μm.

7. The single-cell-capture microfluidic chip of claim 1, wherein: the diameter of the microfluid branch is 10-30 μm.