Polar microorganism separation and culture method based on droplet microfluidic technology

Abstract

The application discloses a polar microorganism separation and culture method based on a droplet microfluidic technology. The method comprises the following steps: obtaining a polar sample and performing oscillation extraction at low temperature to obtain a sample suspension; determining a plate cultivable rate R through a solid plate culture method; based on the R and a preset target number of cultivable cells N, calculating a target total number of cells T required by a droplet microfluidic experiment, and preparing a sample with a corresponding cell concentration for machine loading; generating single-cell-wrapped microdroplets through a droplet microfluidic system for the sample for machine loading, and performing long-term culture at low temperature; and screening positive droplets and purifying strains after the culture. Through the cultivable rate feedback, the sample concentration for the microfluidic loading is precisely optimized, the separation efficiency of the polar difficult-to-culture microorganism is significantly improved, the inter-species competition inhibition of the traditional plate is broken, and the diversity and novel group proportion of the obtained strains are greatly improved.

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