Ultra-sensitive digital quantification of proteins and mRNA in single cells.
Animals
Chlorocebus aethiops
Gene Dosage
Humans
Intravital Microscopy
/ instrumentation
Lab-On-A-Chip Devices
Limit of Detection
Microfluidics
/ instrumentation
Proteins
/ isolation & purification
RNA, Messenger
/ isolation & purification
Single-Cell Analysis
/ instrumentation
Time-Lapse Imaging
/ instrumentation
Vero Cells
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
07 08 2019
07 08 2019
Historique:
received:
29
09
2018
accepted:
16
07
2019
entrez:
9
8
2019
pubmed:
9
8
2019
medline:
18
12
2019
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 31391463
doi: 10.1038/s41467-019-11531-z
pii: 10.1038/s41467-019-11531-z
pmc: PMC6685952
doi:
Substances chimiques
Proteins
0
RNA, Messenger
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Video-Audio Media
Langues
eng
Sous-ensembles de citation
IM
Pagination
3544Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM127527
Pays : United States
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