Ultra-sensitive digital quantification of proteins and mRNA in single cells

Nat Commun. 2019 Aug 7;10(1):3544. doi: 10.1038/s41467-019-11531-z.

Abstract

Simultaneous measurement of proteins and mRNA in single cells enables quantitative understanding and modeling of cellular functions. Here, we present an automated microfluidic system for multi-parameter and ultra-sensitive protein/mRNA measurements in single cells. Our technology improves the sensitivity of digital proximity ligation assay by up to 55-fold, with a detection limit of 2277 proteins per cell and with detection efficiency of as few as 29 protein molecules. Our measurements using this system reveal higher mRNA/protein correlation in single mammalian cells than previous estimates. Furthermore, time-lapse imaging of herpes simplex virus 1 infected epithelial cells enabled by our device shows that expression of ICP4 -a major transcription factor regulating hundreds of viral genes- is only partially correlated with viral protein counts, suggesting that many cells go through abortive infection. These results highlight the importance of high-sensitivity protein/mRNA quantification for understanding fundamental molecular mechanisms in individual cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Chlorocebus aethiops
  • Gene Dosage
  • Humans
  • Intravital Microscopy / instrumentation
  • Intravital Microscopy / methods
  • Lab-On-A-Chip Devices
  • Limit of Detection
  • Microfluidics / instrumentation
  • Microfluidics / methods
  • Proteins / isolation & purification*
  • RNA, Messenger / isolation & purification*
  • Single-Cell Analysis / instrumentation
  • Single-Cell Analysis / methods*
  • Time-Lapse Imaging / instrumentation
  • Time-Lapse Imaging / methods
  • Vero Cells

Substances

  • Proteins
  • RNA, Messenger