Single- and two-photon fluorescence recovery after photobleaching.
Single- and two-photon fluorescence recovery after photobleaching.
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DOI：
10.1101/pdb.top083519
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发表时间：
2015-01-05
影响因子：
--
通讯作者：
Brown EB
中科院分区：
文献类型：
--
作者：
Sullivan KD;Majewska AK;Brown EB
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 chapter discusses the three basic FRAP methods: traditional FRAP, multi-photon FRAP (MPFRAP), and FRAP with spatial Fourier analysis (SFA-FRAP). Each discussion is accompanied by a description of the appropriate mathematical analysis appropriate for situations in which the recovery kinetics are dictated by free diffusion. In some experiments, the recovery kinetics are dictated by the boundary conditions of the system, and FRAP is then used to quantify the connectivity of various compartments. Since the appropriate mathematical analysis is independent of the bleaching method, the analysis of compartmental connectivity is discussed last, in a separate section.
