Single- and two-photon fluorescence recovery after photobleaching.
Cold Spring Harb Protoc
; 2015(1): pdb.top083519, 2015 Jan 05.
Article
in En
| MEDLINE
| ID: mdl-25561627
ABSTRACT
Fluorescence recovery after photobleaching (FRAP) is a microscopy technique for measuring the kinetics of fluorescently labeled molecules and can be applied both in vitro and in vivo for two- and three-dimensional systems. This introduction discusses the three basic FRAP methods:
traditional FRAP, multiphoton FRAP (MPFRAP), and FRAP with spatial Fourier analysis (SFA-FRAP). Each discussion is accompanied by a description of the mathematical analysis appropriate for situations in which the recovery kinetics is dictated by free diffusion. In some experiments, the recovery kinetics is dictated by the boundary conditions of the system, and FRAP is then used to quantify the connectivity of various compartments. Because the appropriate mathematical analysis is independent of the bleaching method, the analysis of compartmental connectivity is discussed last, in a separate section.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Photons
/
Fluorescence Recovery After Photobleaching
/
Photobleaching
Limits:
Animals
/
Humans
Language:
En
Journal:
Cold Spring Harb Protoc
Year:
2015
Document type:
Article