Gradient micro-droplet array forming method based on sequential injection and microfluidic technology

Abstract

The invention discloses a gradient micro-droplet array forming method based on sequential injection and microfluidic technology. The gradient micro-droplet array forming method comprises following steps: step 1, one of a diluent and a sample I is taken using a capillary tube; step 2, the other one of the diluent and the sample I is taken using the capillary tube, and direct contact of the diluent with the sample I is ensured so as to form a sample zone belt with axial concentration gradient; and step 3, the sample zone belt is injected into certain areas of a microporous array chip via the capillary tube so as to obtain sample droplet array with different concentration on the microporous array chip. Advantages of the gradient micro-droplet array forming method are that: concentration gradient generation throughput is high, concentration information is abundant, automatic degree is high; sample consumption is low, and devices and principles are simple. The gradient micro-droplet array forming method can be used for biochemical analysis and screening such as high throughput drug screening, compound toxicity determination, protein crystallization condition screening, catalyst screening, and enzyme dynamic analysis.

Description

technical field [0001] The invention relates to the field of microfluidic droplet analysis and screening, in particular to a method for forming a gradient microdroplet array based on sequential injection and microfluidic droplet technology. Background technique [0002] High Throughput Screening (HTS) uses computer and automated robot technology to complete the complex liquid manipulation, high-sensitivity detection and data processing required for biochemical analysis, and can achieve an analysis throughput of more than 10,000 samples / day. Due to its powerful screening and analysis functions, the application range of high-throughput screening technology has expanded from new drug research and development to biology, medicine, chemistry and other scientific fields, such as structural biology, combinatorial chemistry, synthetic biology, clinical screening As well as occasions where large-scale biochemical analysis experiments are required in various omics studies, it has beco...

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Problems solved by technology

However, the above-mentioned methods all have relatively large limitations, which are mainly reflected in: 1) The continuous flow system is more suitable for screening systems at the molecular level, such as enzyme inhibition screening, etc., but it is difficult to apply to screening systems that require static culture, such as cell Horizontal screening and screening of protein crystallization conditions; 2) The detection of continuous flow systems mainly relies on end-point detection methods such as laser-induced fluorescence, which limits its application in screening systems that require real-time monitoring and imaging analysis; 3) In current systems, enzymes and Reagent solutions such as substrates are continuously injected into the reaction channel, forming a large number of empty reaction droplets, resulting in additional waste of samples; 4) In the continuous flow droplet system, the formation of concentration gradients and the generation of droplets are easily affected by liquid viscosity, temperature Influenced by other factors, the reproducibility of the screening system is not high

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