Drop-based microfluidics (DBM) for single cell studies and ultrahigh-throughput phenotypic screens and lab evolution

点击翻译按钮获取中文摘要

There is heterogeneity in how individual cells in a population process information and respond to both genetic and environmental perturbations. Traditional bulk assays with readouts averaged over all cells in the population may not reflect how individual cells behave. It is therefore essential to be able to isolate, manipulate, and perform analysis at the level of single cells to capture the full diversity of biological processes, including those that are rare in the population. ******Multiple research groups in the Departments of Biochemistry and Biology at Université de Montréal (UdeM) are requesting a custom-made Drop-Based Microfluidics (DBM) for single cell studies and ultrahigh-throughput phenotypic screens and lab evolution. The research groups of the five applicants, together with their collaborators, study diverse and fundamental aspects of biology, such as structure-function relationship of complex macromolecules, and whole-cell interactome and genomics, as well as microbial evolution and population dynamics. However, they have a common goal of dissecting at the level of single cell the population-level heterogeneity of these processes. The DBM will be a crucial tool for the success of their NSERC funded research programs. It will also provide training for students and postdocs to acquire high-level skills that are important for both the biotech and pharmaceutical sectors.******At its core, the DBM uses picoliter droplet emulsions numbering ~106 up to ~108 as vessels or “test-tubes” for assays, isolation of cells, preparation of genomic libraries for next-generation sequencing, and for serial propagation of cultures for lab evolution. DBM allows up to ~108 samples to be screened in one day. Compartmentalization in droplets dramatically improves assay sensitivity by increasing the effective concentration of reactants and decreasing the time required to reach detection thresholds. Because of these capabilities, DBM has been used for preparing libraries of biological samples for single-cell transcriptomics, Chipseq, and DNAseq; for isolation, detection and enrichment of rare genomic events; and for high-throughput screening and microbial lab evolution. There is currently no such DBM set-up in the Biochemistry Department nor in any other departments and institutes in Quebec. Thus its purchase is urgently needed to perform state-of-the-art single cell experiments.