Details

Autor(en) / Beteiligte

• Lin, Jing
• Jordi, Christian
• Son, Minjun
• Van Phan, Hoang
• Drayman, Nir
• Abasiyanik, Mustafa Fatih
• Vistain, Luke
• Tu, Hsiung-Lin
• Tay, Savaş

Titel

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

Ist Teil von

• Nature communications, 2019-08, Vol.10 (1), p.3544-10, Article 3544

Ort / Verlag

England: Nature Publishing Group

Erscheinungsjahr

2019

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.