CHAPTER 5: Microfluidic Applications in Single-cell Genomic, Transcriptomic and Proteomic Analysis
Published:20 Nov 2020
Special Collection: 2020 ebook collectionSeries: Soft Matter Series
D. Dhingra, A. Ooi, P. Mendez, S. Wang, S. Gulati, A. Sciambi, and D. Ruff, in Droplet Microfluidics, ed. C. Ren and A. Lee, The Royal Society of Chemistry, 2020, pp. 122-146.
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Advances in microfluidic droplet technologies have greatly facilitated breakthroughs in cellular and biomedical sciences, particularly in the arena of single-cell genomic, transcriptomic and proteomic analysis applications. Microfluidic devices enable the high-throughput preparation of biological samples that are readily partitioned into nanoliter-scale droplets containing isolated individual cells or nuclei. Aqueous droplets are suspended and isolated in a hydrophobic carrier liquid. Typical microfluidic workflows offer the magnitude of multiple thousands upward to millions of encapsulated droplets per experimental run. Biochemical reactions provide for the ready interrogation of cellular components such as genomic DNA, RNA transcripts and protein markers. Innovative tagging and nucleic acid amplification workflows generate droplet-specific barcode identifiers compatible with next-generation sequencing platforms. After barcoding, thousands of droplets are combined together, amplified offline, size purified and sequenced in massive parallel protocols. One such enabling platform is the Mission Bio Tapestri® system. This unique system offers a novel two-stage droplet workflow that provides multiple sequential cascades of enzymatic and chemistry processes, and opens opportunities for multimodal investigations of single cells. We describe Tapestri applications including workflows, reaction overviews and data examples. These microfluidic applications have propelled the frontiers of cellular analysis to further multidimensional analysis of cell velocities in dynamic biological systems.