Agarose Beads-based Long-term Single-cell Culture And Sorting For Rapid Growth Haematococcus Pluvialis Microalgae Screening

Microfluidic technology for microbial strain culture and screening can effectively solve above mentioned problems. Microfluidics or lab on a chip technology has been introduced in analytical chemistry since 1990s. It has been widely used because of its high integration, real-time monitoring and precise manipulation of samples from pL to μL, which potentially provides a strong technical support for high-throughput strains screening. The development of single cell screening method may greatly improve the screening efficiency and shorten the screening period compared to traditional strain screening methods. Each microspheres can be accurately controlled through chip design and external controlling. In this study, microalgae haematococcus pluvialis was used as a model. And a platform for screening of microbial strain with high growth rate using droplet droplet microfluidics technology has been developed.1) single cell encapsulation and cultureBy using water in oil gel microspheres technology and hydrodynamic droplet encapsulation method, single microalgae cells for culture. A high throughput screening based on the optical density difference originated from the cell growth difference encapsulated in different agaroses beads was developed. The whole process was describe as follows. Firstly, single microalgae cell was encapsulate into agarose droplet on the microfluidics chip. After solidification on the ice bath and oil degreasing, the prepared beads were re-suspended in 3N BBM medium to incubate for 4 to 7 days.2) microgel screening for Rapid Growth Haematococcus pluvialisThe cultured agarose beads were re-injected into microfluidics chip for sorting. The online optical detector and sorting equipment were developed to distinguish and sort the agarose beads with bigger haematococcus pluvialis microalgae cell which would induce higher optical density signal because of bigger biomass for cell with higher growth rate. Microfluidic technology usingagarose beads for single cell culture and screening can accurately identify and separate strains at the single cell level. Compared with the traditional microbial cultivation and screening methods, the developed platform holds several significant advantages of was high throughput, high automation, low reagent consumption and time-saving which laid the technologies and methods foundation for high throughput industrial microbial screening in the near future.