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

摘要: Simultaneous measurement of proteins and mRNA in single cells enables quantitative understanding modeling cellular functions. Here, we present an automated microfluidic system for multi-parameter ultra-sensitive protein/mRNA measurements cells. Our technology improves the sensitivity digital proximity ligation assay by up to 55-fold, with a detection limit 2277 per cell efficiency as few 29 protein molecules. using this reveal higher mRNA/protein correlation mammalian than previous estimates. Furthermore, time-lapse imaging herpes simplex virus 1 infected epithelial enabled our device shows that expression ICP4 -a major transcription factor regulating hundreds viral genes- is only partially correlated counts, suggesting many go through abortive infection. These results highlight importance high-sensitivity quantification fundamental molecular mechanisms individual Digital (dPLA) can measure mRNAs cells, but not compatible cannot quantify rare due high dilution factor. Here authors combines live-cell imaging, chemical stimulation, dPLA smaller reaction volume.