doiSerbia

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

Jovanović-Talisman Tijana (Department of Molecular Medicine, Beckman Research Institute of the City of Hope Comprehensive Cancer Center, Duarte, USA + Department of Chemistry, University of Hawaii at Manoa, Honolulu, Hawaii, USA)
Vukojević Vladana (Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden)

Self-organization of cell-surface receptors in structurally distinct domains in the plasma membrane is of vital interest for correct cellular signaling. However, this dynamic process is difficult to study in cells with sufficiently high temporal and spatial resolution. We present here two quantitative high-resolution methods with single-molecule sensitivity, Fluorescence Correlation Spectroscopy (FCS) and pair-correlation Photoactivated Localization Microscopy (pcPALM), which enable nondestructive study of receptor diffusion and lateral organization at the nanoscale level. We introduce here the methods and review their application in studies of lateral organization of G Protein-Coupled Receptors (GPCRs). Examples from our own work on opioid receptor lateral organization are presented in order to illustrate the most recent advances in the field.

Keywords: fluorescence correlation spectroscopy (FCS), super-resolution fluorescence imaging, sub-diffraction limit, photoactivated localization microscopy (PALM), G protein-coupled receptors (GPCRs), lipid rafts

Projekat Ministarstva nauke Republike Srbije, br. 172015 i br. 45001