Scientists develop high-precision droplet printing platform for single cell phenotype screening

With the speedy improvement of genome synthesis and enhancing applied sciences, droplet-based single-cell phenotype screening has change into more and more necessary. Nonetheless, exact and high-throughput distribution of goal droplets into particular macroscopic media for downstream multi-omics evaluation stays a technical problem.

Researchers from the Qingdao Institute of Bioenergy and Bioprocess Know-how (QIBEBT) of the Chinese language Academy of Sciences have developed a droplet-based printing platform that may print single-cell droplets on tradition dishes or 96-well plates in a “one droplet per properly” method. This platform permits scalable, cost-effective, and absolutely automated single-cell phenotype seize and screening.

The work is printed in Sensors and Actuators B: Chemical.

Agar plate and microplate screening are the 2 most generally used conventional screening strategies. Agar plate assays are straightforward to function however lack exact quantification and are subsequently primarily used for preliminary pressure screening. Microplate assays present correct quantification and are appropriate for evaluating mutant efficiency, however have decrease throughput.

Droplet microfluidics–primarily based instruments maintain nice promise for ultra-high throughput screening. Nonetheless, present sorting methods usually solely enrich droplets with a specific phenotype in a single channel, making it difficult to exactly and effectively separate every goal droplet to particular areas inside macroscopic media (e.g., tradition dishes or 96-well plates) for subsequent single-cell tradition and sequencing experiments.

The droplet printing platform developed on this examine consists of 4 predominant modules: A pump-driven microfluidic chip module, an computerized optical sign recognition module, a high-pressure management module, and an automatic cellular platform module.

Diao Zhidian, a Ph.D. scholar and co-first writer of the examine, defined that the chip comprises embedded optical fibers that may introduce pink light-emitting diodes. When the sorted goal droplets go by the detection window, the photodiodes related to the optical fibers can determine them and generate corresponding alerts. These alerts are then used to print the droplets in particular areas primarily based on electrohydrodynamic ideas.

“This chip platform has a number of distinctive options,” mentioned Ge Anle, co-first writer of the examine. “Not solely can this platform generate droplets on-line, however it could additionally precisely print incubated or sorted droplets precisely on demand, changing ‘goal droplet teams’ into ‘particular person goal droplets,’ thereby serving downstream phenotype research on the single cell stage.”

“This platform incorporates an optical detection module primarily based on optical fiber transmission,” mentioned co-corresponding writer Prof. Ma Bo. “This module not solely improves the accuracy of droplet printing, but additionally avoids complicated optical path methods. This concise and built-in system is suitable with a wider vary of downstream experimental eventualities, comparable to mass spectrometry, cell tradition dishes, and 96-well plates.”

Subsequent, the analysis crew plans to combine this high-precision on-demand single-droplet printing platform with the beforehand developed Raman-activated cell sorting devices, comparable to FlowRACS and RACS-Seq/Tradition, in order to realize absolutely automated, high-precision, high-throughput useful single-cell identification, separation, tradition, and export for complicated organic methods comparable to microbiomes and large-scale mutant libraries.

“The mixing of our platform with current single-cell Raman sorting devices comparable to FlowRACS will open up new prospects for large-scale single-cell useful research, offering a complete resolution for biobanking, artificial biology, precision medication, deep-sea/deep-Earth/deep-space exploration, and biosecurity,” mentioned Prof. Xu Jian, head of the Single-Cell Middle of QIBEBT.