Abstract
We introduce Digital microfluidic Isolation of Single Cells for -Omics (DISCO), a platform that allows users to select particular cells of interest from a limited initial sample size and connects single-cell sequencing data to their immunofluorescence-based phenotypes. Specifically, DISCO combines digital microfluidics, laser cell lysis, and artificial intelligence-driven image processing to collect the contents of single cells from heterogeneous populations, followed by analysis of single-cell genomes and transcriptomes by next-generation sequencing, and proteomes by nanoflow liquid chromatography and tandem mass spectrometry. The results described herein confirm the utility of DISCO for sequencing at levels that are equivalent to or enhanced relative to the state of the art, capable of identifying features at the level of single nucleotide variations. The unique levels of selectivity, context, and accountability of DISCO suggest potential utility for deep analysis of any rare cell population with contextual dependencies.
Highlights
We introduce Digital microfluidic Isolation of Single Cells for -Omics (DISCO), a platform that allows users to select particular cells of interest from a limited initial sample size and connects single-cell sequencing data to their immunofluorescence-based phenotypes
DISCO is the first method to mate digital microfluidics (DMF)[27] with laser cell lysis (LCL)[26], a technique in which a focused, highenergy laser is used to lyse cells within a few micrometers of the focal point
DISCO (Figs. 1, 2) is an addition to the canon of microfluidic techniques that have been developed for single-cell -Omics analysis[7,8,9,10,11,12,13,14,15,16,17,18,19,20,21]
Summary
We introduce Digital microfluidic Isolation of Single Cells for -Omics (DISCO), a platform that allows users to select particular cells of interest from a limited initial sample size and connects single-cell sequencing data to their immunofluorescence-based phenotypes. Parker et al.[24] described a creative method relying on photoelectrochemical effects to isolate individual cells from adherent culture prior to sequencing, but the technique requires specialized gridded substrates that force artificial limits on cell spacing. These techniques (and LCM, and mechanical pickers and patterned/ specialty substrates, in general) are important and useful, but we are not aware of a robust, all-purpose path for the user to connect adherent cell phenotype to single-cell genomes, transcriptomes, or proteomes. We propose that DISCO represents an important tool in the single-cell analyst’s toolbox, that should be uniquely well suited for assessing questions related to the fundamental dogma of molecular biology, testing the relationship between genome, transcriptome, and proteome and phenotype
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