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High-throughput nano-litre micro-droplet forming and fixing method based on microfluidic chip and special chip and application thereof

A technology of microfluidic chip and immobilization method, which is applied in the direction of supporting/immobilizing microorganisms, biochemical equipment and methods, measuring/inspecting microorganisms, etc. Advanced problems, to achieve the effect of simplifying the operation process, shortening the experiment time and reducing consumption

Inactive Publication Date: 2009-11-04
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the automated operation speeds up the analysis, there are still many limitations: such as micro-sample manipulation requires high precision of the automated equipment; open environment micro-volume solution is easy to evaporate; sample pretreatment (including nematode culture, drug application, Washing, etc.) process is still offline, resulting in cumbersome analysis process and low throughput; the huge volume and high price of instruments and equipment make it difficult to be widely used, etc.
Overall, the current development of model biotechnology is far from meeting the needs of modern biomedical research and development. Therefore, it is important to develop a new platform for high-throughput drug screening based on model organisms and to realize the miniaturization, integration, and low cost of screening technologies. The inevitable trend of future development

Method used

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  • High-throughput nano-litre micro-droplet forming and fixing method based on microfluidic chip and special chip and application thereof
  • High-throughput nano-litre micro-droplet forming and fixing method based on microfluidic chip and special chip and application thereof
  • High-throughput nano-litre micro-droplet forming and fixing method based on microfluidic chip and special chip and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Will figure 1 The inlets of the continuous phase and the dispersed phase on the microfluidic chip shown are respectively connected to two syringe pumps through Teflon tubes. Driven by the syringe pumps, hexadecane and green ink are respectively injected into the chip. The droplet generation zone forms continuous green microdroplets (see Figure 4 a); the micro-droplets continue to flow into the droplet capture array driven by the syringe pump, and are sequentially captured in the circular droplet catcher (see Figure 4 b); Finally, the syringe pump is turned off to stop the droplet generation and fluid flow, to obtain immobilized uniform microdroplets of 180 nanoliters.

Embodiment 2

[0046] Will figure 1 The inlets of the continuous phase and the dispersed phase on the microfluidic chip shown are respectively connected to two syringe pumps through Teflon tubes. Driven by the syringe pumps, hexadecane and nematode suspensions are respectively injected into the chip. The microdroplet generation zone forms continuous microdroplets encapsulating individual nematodes (see Figure 5 a); the micro-droplets continue to flow driven by the syringe pump, enter the droplet capture array and are sequentially captured in the circular droplet catcher (see Figure 5 b, 6a); turn off the syringe pump to stop the droplet production and fluid flow, place the microfluidic chip under a stereo microscope, and analyze the movement behavior of a single nematode in the droplet microenvironment (see Figure 7-Figure 10 ). From the results, it can be found that within the detection time (2 hours), the movement of nematodes in the droplet maintains a high oscillation frequency (abo...

Embodiment 3

[0048] Will figure 1 The inlets of the continuous phase and the dispersed phase on the microfluidic chip shown are respectively connected to two syringe pumps through Teflon tubes. Driven by the syringe pumps, hexadecane and MPP+ containing a certain concentration of neurotoxins (3mM, 5mM ) of the nematode suspension were injected into the chip respectively, and a continuous micro-droplet wrapped with a single nematode was formed in the T-shaped micro-droplet generating area (see Figure 5 a); the micro-droplets continue to flow driven by the syringe pump, enter the droplet capture array and are sequentially captured in the circular droplet catcher (see Figure 5 b, 6a); turn off the syringe pump to stop the droplet production and fluid flow, place the microfluidic chip under a stereo microscope, and detect the locomotor behavior (oscillating frequency, Ω shape) of a single nematode under the action of different concentrations of neurotoxin MPP+ frequency of occurrence). Fi...

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Abstract

The invention relates to a high-throughput nano-litre micro-droplet forming and fixing method based on a microfluidic chip and a special chip and an application thereof. The method comprises the following steps: inlets of a continuous phase and a dispersed phase on the chip are respectively connected with two injection pumps through a teflon tube, the continuous phase and the dispersed phase are respectively infused into the chip under the driving action of the injection pumps, and continuous micro-droplets wrapped with single nematodes are formed in a T-shaped micro-droplet generation area; the generated micro-droplets enter in a droplet capture array and are captured in a circular droplet catcher in sequence; and after the micro-droplets are fully captured, the behaviors of the single nematodes in the droplet microenvironment are detected. The invention has the advantages that the microfluidic chip droplet technology is combined with the behavioral analysis to the single nematodes, a great amount of micro-droplets wrapped with the single nematodes are generated and fixed within the short time by using the characteristics of flexible combination and large-scale integration of various functional units on the chip, and the behavioral analysis to the high-throughput single nematodes on the microfluidic chip is realized for the first time.

Description

technical field [0001] The present invention relates to a droplet formation and immobilization method based on a microfluidic chip and its application. In particular, it provides a high-throughput nanoscale microdroplet formation and immobilization method based on a microfluidic chip, its dedicated chip, and the behavior of a single nematode. applications in analysis. Background technique [0002] Microfluidic chip (microfluidic chip), also known as lab on a chip, refers to the integration or basic integration of basic operating units such as sample preparation, reaction, separation, and detection involved in the fields of biology and chemistry. On a chip of a few square centimeters (or even smaller), a network of microchannels is formed, and a controllable fluid runs through the entire system to replace various functions of conventional biological or chemical laboratories. Its basic feature and greatest advantage are the flexible combination and large-scale integration of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/02C12M1/00
CPCC12M23/16C12M25/01
Inventor 秦建华施维维叶囡楠林炳承
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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