The Utility of Fluorescence Recovery after Photobleaching (FRAP) to Study the Plasma Membrane.

Content Provider	Europe PMC
Author	Day, Charles A. Kang, Minchul
Editor	Urbančič, Iztok Schneider, Falk
Copyright Year	2023
Abstract	The plasma membrane of mammalian cells is involved in a wide variety of cellular processes, including, but not limited to, endocytosis and exocytosis, adhesion and migration, and signaling. The regulation of these processes requires the plasma membrane to be highly organized and dynamic. Much of the plasma membrane organization exists at temporal and spatial scales that cannot be directly observed with fluorescence microscopy. Therefore, approaches that report on the membrane’s physical parameters must often be utilized to infer membrane organization. As discussed here, diffusion measurements are one such approach that has allowed researchers to understand the subresolution organization of the plasma membrane. Fluorescence recovery after photobleaching (or FRAP) is the most widely accessible method for measuring diffusion in a living cell and has proven to be a powerful tool in cell biology research. Here, we discuss the theoretical underpinnings that allow diffusion measurements to be used in elucidating the organization of the plasma membrane. We also discuss the basic FRAP methodology and the mathematical approaches for deriving quantitative measurements from FRAP recovery curves. FRAP is one of many methods used to measure diffusion in live cell membranes; thus, we compare FRAP with two other popular methods: fluorescence correlation microscopy and single-particle tracking. Lastly, we discuss various plasma membrane organization models developed and tested using diffusion measurements.
Volume Number	13
PubMed Central reference number	PMC10221041
Issue Number	5
PubMed reference number	37233553
Journal	Membranes (Basel)
e-ISSN	20770375
DOI	10.3390/membranes13050492
Language	English
Publisher	MDPI
Publisher Date	2023-05-02
Access Restriction	Open
Rights License	Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). © 2023 by the authors.
Subject Keyword	fluorescence recovery after photobleaching FRAP diffusion plasma membrane microdomain actin confocal microscopy
Content Type	Text
Resource Type	Article
Subject	Chemical Engineering Process Chemistry and Technology Filtration and Separation