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Micro-fluidic chip for accurately assembling single particles and single particle assembling method

A microfluidic chip and particle technology, applied in laboratory containers, chemical instruments and methods, laboratory utensils, etc., can solve the problem of inability to achieve precise assembly, capture and assembly of difficult types of single particles, and inability to accurately control the number of cells And other issues

Active Publication Date: 2021-07-06
厦门德运芯准科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the controllable assembly capabilities of existing microfluidic chip devices are limited
For example, microarrays or droplet microfluidic systems are limited by the Poisson distribution, resulting in most micropores or droplets not containing single particles, and only a few containing single particles, which is even less accurate. Control the number of cells assembled in microwells or droplets
Most other types of microfluidic devices, such as microfluidic chips based on hydrodynamic trapping principles, still cannot achieve true precise assembly
In addition, in recent years, some microfluidic chips that can be used for the assembly of multiple particles have emerged. One operation can only achieve the capture and assembly of one type of single particle, and it is difficult to capture two different types of single particles at the same time in a single operation. and assemble

Method used

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  • Micro-fluidic chip for accurately assembling single particles and single particle assembling method
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  • Micro-fluidic chip for accurately assembling single particles and single particle assembling method

Examples

Experimental program
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Effect test

Embodiment 1

[0055] Example 1 PDMS microfluidic chip for precise assembly of single particles

[0056] Chip reference of the present invention figure 1 As shown, the chip structure consists of two parts, the channel layer A and the sealing layer B, wherein the channel layer is located on the upper layer, and the sealing layer is located on the lower layer. The bottom of the channel layer A is provided with grooves to form the capture channel 1 and the assembly cavity 2, wherein, The height of the assembly chamber 2 is higher than that of the capture channel 1 .

[0057] The channel structure includes a plurality of basic units connected together. In this embodiment, these basic units are connected in series (such as figure 2 and image 3 shown). The connection mode of the basic units is not limited to the method in this embodiment, and can also be connected in parallel.

[0058] Chip channel layer structure reference Figure 1 to Figure 3 , including 2 channels (i.e. the first channe...

Embodiment 2

[0066] Example 2 Precise assembly of single particles

[0067] As a preferred embodiment of the present invention, the cells used are K562 cells with a diameter of 10-20 microns, and the two kinds of microspheres used are polystyrene microspheres with a diameter of 20 microns and 40 microns.

[0068] As a preferred embodiment of the present invention, the selected cell flow rate is 0.02ml / h. As a preferred embodiment of the present invention, the flow rate of the selected microspheres is 0.05ml / h.

[0069] As a preferred embodiment of the present invention, the flow rate of backflushing is 0.05ml / h.

[0070] The specific working process of accurately assembling single particles is as follows:

[0071] Step A: Perfuse the solution from the two parallel channels of the chip separately to eliminate air bubbles on the chip, and at the same time seal the chip to prevent the adhesion of single particles. Next, the solution is replaced with a single-particle suspension, and sample...

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Abstract

The invention discloses a micro-fluidic chip for accurately assembling a single particle and a single particle assembling method. The micro-fluidic chip structurally comprises a channel layer and a sealing layer, wherein the channel layer structurally comprises two parallel and independent channels, and each channel comprises a capture channel, an annular bypass and an assembly chamber. The method comprises the following operation steps: (1) capturing a single particle; (2) transferring the single particle to an assembly cavity by reversely introducing a solution; and (3) capturing and transferring multiple times to realize multi-cell assembly. According to the invention, tThe chip can achieve efficient single cell capture based on the fluid mechanics principle, assembly of single particles is controlled by controlling the flow direction of fluid, and accurate assembly of the single particles can be achieved through multiple times of repeated capture and assembly; and the micro-fluidic chip can be applied to the research and application fields of single-cell RNA sequencing, single-cell co-culture, cell secreted protein detection, single-cell interaction analysis and the like.

Description

technical field [0001] The invention relates to a microfluidic chip for precisely assembling single particles, which can be applied to fields such as single-cell RNA sequencing, single-cell co-cultivation, detection of cell secreted proteins, single-cell interaction analysis, and the like. Background technique [0002] The assembly of single particles is of great significance for studying the properties and functions of particles. Taking single cells as an example, cells have strong heterogeneity. Traditional analysis based on population cells often conceals the heterogeneity between cells and obtains the average expression Information and, in many cases, individualized single-cell functions play an important role in physiology as well as disease. Microfluidic chip is called "lab-on-a-chip", which has the advantages of miniaturization, integration, high throughput, size and single particle matching, so it is especially suitable for the analysis of single particle. However, ...

Claims

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Application Information

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IPC IPC(8): B01L3/00
CPCB01L3/5027B01L3/502761B01L2300/0861B01L2300/0819B01L2300/0681
Inventor 杨朝勇黄山青刘卫枝朱志周雷激
Owner 厦门德运芯准科技有限公司
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