Ultra-high throughput single-cell analysis of proteins and RNAs by split-pool synthesis

Commun Biol. 2020 May 7;3(1):213. doi: 10.1038/s42003-020-0896-2.

Abstract

Single-cell omics provide insight into cellular heterogeneity and function. Recent technological advances have accelerated single-cell analyses, but workflows remain expensive and complex. We present a method enabling simultaneous, ultra-high throughput single-cell barcoding of millions of cells for targeted analysis of proteins and RNAs. Quantum barcoding (QBC) avoids isolation of single cells by building cell-specific oligo barcodes dynamically within each cell. With minimal instrumentation (four 96-well plates and a multichannel pipette), cell-specific codes are added to each tagged molecule within cells through sequential rounds of classical split-pool synthesis. Here we show the utility of this technology in mouse and human model systems for as many as 50 antibodies to targeted proteins and, separately, >70 targeted RNA regions. We demonstrate that this method can be applied to multi-modal protein and RNA analyses. It can be scaled by expansion of the split-pool process and effectively renders sequencing instruments as versatile multi-parameter flow cytometers.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / analysis*
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Proteins / analysis*
  • RNA / analysis*
  • Single-Cell Analysis / methods*

Substances

  • Antibodies
  • Proteins
  • RNA