Laser fluorescence photobleaching recovery mobility

Fluorescence microphotolysis, or photobleaching, has been widely used to study translational mobility of lipids and proteins in membranes. An attenuated laser beam may be focused down to the diameter of a cell or less. Then the intensity can be suddenly increased by several orders of magnitude, bleaching any fluorescent material present. The return of fluorescent material by free diffusion from a neighboring region (fluorescence recovery after photobleaching) or by diffusion through a membrane into a cell can then be... 

This chapter reviews several techniques which combine the use of laser microbeams with antibodies to study molecular and cellular biology. An overview of the basic properties of lasers and their integration with microscopes and computers is provided. Biophysical applications, such as fluorescence recovery after photobleaching to measure molecular mobility and fluorescence resonance energy transfer to measure molecular distances, as well as ablative applications for the selective inactivation of proteins or the selective killing of cells are described. Other techniques, such as optical trapping, that do not rely on the interaction of the laser with the targeting antibody, are also discussed. 

Braga, J. Desterro, J. M. et al. (2004). "Intracellular macromolecular mobility measured by fluorescence recovery after photobleaching with confocal laser scanning microscopes." Mol Biol Cell, 15(10), 4749-60. 

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