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Fluorescence bleach rate imaging
Authors:G J BRAKENHOFF  K VISSCHER  E J GIJSBERS
Abstract:Bleach rate imaging on a (cooled) CCD can be easily achieved using a confocal microscope with bilateral scanning and detection coupled to a workstation; it is as easy as acquiring regular fluorescence images. Several analysis and display methods for bleach rate imaging are presented such as the bleach map (and its inverse) and a matrix-based decomposition method for multi-labelled specimens based on the bleach rate differences between the dyes used. With these tools, bleach-rate-based imaging can become a viable alternative to multiple labelling techniques for component identification in fluorescent specimens.
Keywords:Fluorescence microscopy  photobleaching  bleach rate  bleach map  histomap  confocal  bilateral  ratio-imaging  CCD  photodegradation
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