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Method of forming and separately exporting single particle wrapped drop in micro-fluidic chip

A microfluidic chip and particle technology, applied in the field of microfluidics, can solve the problems of low throughput, short detection time, slow progress, etc., and achieve the effects of low impact on cell activity, reduced operating costs, and fast pipetting speed

Active Publication Date: 2019-05-21
青岛星赛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the progress in single cell capture and separation and extraction has been slow. Through microfluidic chip technology, it can be roughly divided into two methods: single cell array capture and flow cytometry.
The method of array capture can realize real-time monitoring of single cells, and it is convenient to change the experimental conditions and study the influence of different culture conditions on single cells, but the throughput is low
Flow cytometry methods are usually based on single-cell droplet packaging technology, which has the advantages of high detection throughput and convenient cell separation. However, due to the short detection time, it is generally difficult to obtain more detailed information on a single cell. host
However, how to separate and take out the cells analyzed and screened in the microfluidic chip, and then perform downstream analysis is still a current difficulty

Method used

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  • Method of forming and separately exporting single particle wrapped drop in micro-fluidic chip
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  • Method of forming and separately exporting single particle wrapped drop in micro-fluidic chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] 1. Preparation of microfluidic chip:

[0063] ①Use the method of ultrasonic drilling to punch the sample injection hole and the reservoir hole on the upper quartz glass, and the hole spacing is according to the channel length. The upper layer of quartz glass is a smooth plane with a thickness of 0.5-1 mm and no etching pattern. (The sequence of the upper and lower layers of quartz glass depends on the optical path of the optical tweezers. Since the microscope used in this experiment is an upright microscope, the optical path of the optical tweezers passes through the chip from top to bottom, so it is necessary to ensure that the upper surface of the chip is a smooth optical surface, so the upper layer is smooth. Quartz glass, the quartz glass with engraved channels is on the lower level)

[0064] ②The surface of the lower quartz glass is etched according to the channel design, and the channel height is about 15-50 microns.

[0065] ③The two layers of glass are aligned...

Embodiment 2

[0072] The formation and export of the target single particle-encapsulated droplet, the schematic diagram of the steps is shown in Figure 4 .

[0073] ①Inject the oil phase into the oil reservoir at the outlet end of the sample channel (generally, mineral oil containing surfactant is used).

[0074] ②Using the method of static pressure injection, the particle phase (cell phase is used in this example) solution is injected into the channel of the chip through the injection port, and the height of the sample holder is adjusted to h0 so that the interface between the cell phase and the oil phase of the sample outlet Stable and stationary near the mouth.

[0075] ③ Collect the characteristic map of a single cell in the detection cell (such as Raman spectrum, 532nm laser), and capture the desired target cells through optical tweezers (such as 1064nm laser), move the microfluidic chip, and drag the target cells to the water phase Near the interface with the oil phase.

[0076] ④...

Embodiment 3

[0080] DNA amplification and electrophoretic detection of isolated single cells.

[0081] The liquid droplets separated in Example 2 are observed under the 50X objective lens, and it can be seen that single cells are successfully wrapped in the liquid droplets. This method is applicable to cells of different sizes, from about one micron to tens of microns, such as Figure 5 G and Figure 5 H is the encapsulation of Escherichia coli cells and yeast cells in the droplet using this method, respectively. Since the density of the mineral oil used in this method is lower than that of the cell suspension (water), the droplets will be automatically distributed at the bottom of the capillary during the transfer of the droplets, and it is only necessary to touch the capillary containing the droplets to the wall of the test tube or centrifuge tube. Droplets can be exported, which makes the transfer process more successful and simpler.

[0082] Centrifuge the single-cell-encapsulated dr...

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Abstract

The invention provides a micro-fluidic chip capable of being used for single particle screening and forming drop wrappage for exporting. The micro-fluidic chip comprises at least one sample channel and at least one oil reservoir. The micro-fluidic chip is connected with a liquid sample introduction device to form a micro-fluidic chip device for forming a single particle wrapped drop, and the micro-fluidic chip device and a particle capture device can further form a microfluidic operating system capable of being used for forming the single particle wrapped drop. The invention further provides amethod of forming and separately exporting the target single particle wrapped drop in the micro-fluidic chip.

Description

technical field [0001] The present invention relates to the field of microfluidic technology, in particular to a method for using a microfluidic chip to form a single particle-wrapped droplet and derive it, which can be used for single-cell screening, single-cell separation, single-cell sequencing, single-cell morphology analysis, single-cell cultivation, drug screening and other fields. Background technique [0002] It is well known that cell diversity is ubiquitous in various cell populations, such as bacteria, yeast, mammalian cells, etc. Even daughter cells that are derived from the same mother cell can vary among individual cells. Single-cell analysis (single-cell analysis) refers to the research on the differences between individual cells in the cell population, including cell size, growth rate, chemical composition (phospholipids, proteins, metabolites, DNA / RNA), and other aspects, as well as cell The reason and mechanism of the difference. Its research content inv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01L3/00
CPCB01L3/502761B01L3/502707B01L2300/0887B01L2300/0883B01L2300/0816B01L2200/0668B01L2200/027B01L2200/0673B01L2400/043B01L2400/0454B01L2300/165B01L2400/0457C12M47/04B01L3/502715B01L2200/0652B01L2300/08B01L2400/0406
Inventor 徐腾马波徐健
Owner 青岛星赛生物科技有限公司
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