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

Nature Communications
Jing LinSavaş Tay

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.

References

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Citations

May 24, 2020·Cells·Lixing LiuJian Chen
May 27, 2020·Cell Communication and Signaling : CCS·Christina N Byrne-HoffmanDavid J Klinke
Mar 4, 2021·Trends in Biochemical Sciences·Luke F Vistain, Savaş Tay
Jul 13, 2021·Analytical Chemistry·Qingxuan LiMing Su

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Methods Mentioned

BETA
enzyme-linked immunosorbent assay
ELISA
proximity ligation assay
PCR
chip
biopsy
chips
Assay

Software Mentioned

Matlab
Elements AR
ImageJ
QuantaSoft
dPLA
NIS

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