Super-resolution fluorescence imaging and correlation spectroscopy: Principles and examples of application

Abstract

Self-organization of cell-surface receptors in structurally distinct domains in the plasma membrane is of vital importance for correct cellular signaling. However, this dynamic process is difficult to study in cells with sufficiently high temporal and spatial resolution. Herein, two quantitative highresolution methods with single-molecule sensitivity are presented, i.e., fluorescence correlation spectroscopy (FCS) and pair-correlation photo-activated localization microscopy (pcPALM), which enable the non-destructive study of receptor diffusion and lateral organization at the nanoscale level. The methods are introduced and their application in studies of lateral organization of G protein-coupled receptors (GPCRs) is reviewed. Examples from studies on the lateral organization of opioid receptors are presented in order to illustrate the most recent advances in the field. ©2013 SCS.