Quantitative mapping of fluorescently tagged cellular proteins using FCS-calibrated four-dimensional imaging.

ABSTRACT: The ability to tag a protein at its endogenous locus with a fluorescent protein (FP) enables quantitative understanding of protein dynamics at the physiological level. Genome-editing technology has now made this powerful approach routinely applicable to mammalian cells and many other model systems, thereby opening up the possibility to systematically and quantitatively map the cellular proteome in four dimensions. 3D time-lapse confocal microscopy (4D imaging) is an essential tool for investigating spatial and temporal protein dynamics; however, it lacks the required quantitative power to make the kind of absolute and comparable measurements required for systems analysis. In contrast, fluorescence correlation spectroscopy (FCS) provides quantitative proteomic and biophysical parameters suc

SUBMITTER: Politi AZ 

PROVIDER: S-EPMC6609853 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature