Quantifying spatial organization in point-localization superresolution images using pair correlation analysis.

ABSTRACT: The distinctive distributions of proteins within subcellular compartments both at steady state and during signaling events have essential roles in cell function. Here we describe a method for delineating the complex arrangement of proteins within subcellular structures visualized using point-localization superresolution (PL-SR) imaging. The approach, called pair correlation photoactivated localization microscopy (PC-PALM), uses a pair-correlation algorithm to precisely identify single molecules in PL-SR imaging data sets, and it is used to decipher quantitative features of protein organization within subcellular compartments, including the existence of protein clusters and the size, density and number of proteins in these clusters. We provide a step-by-step protocol for PC-PALM, illustrati

SUBMITTER: Sengupta P 

PROVIDER: S-EPMC3925398 | biostudies-literature | 2013 Feb

REPOSITORIES: biostudies-literature