Journal
FEBS LETTERS
Volume 546, Issue 1, Pages 87-92Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S0014-5793(03)00521-0
Keywords
fluorescent protein; multicolor microscopy; fluorescence resonance energy transfer; live cell imaging; image analysis
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In biological microscopy, the ever expanding range of applications requires quantitative approaches that analyze several distinct fluorescent molecules at the same time in the same sample. However, the spectral properties of the fluorescent proteins and dyes presently available set an upper limit to the number of molecules that can be detected simultaneously with common microscopy methods. Spectral imaging and linear un-mixing extends the possibilities to discriminate distinct fluorophores with highly overlapping emission spectra and thus the possibilities of multicolor imaging. This method also offers advantages for fast multicolor time-lapse microscopy and fluorescence resonance energy transfer measurements in living samples. Here we discuss recent progress on the technical implementation of the method, its limitations and applications to the imaging of biological samples. (C) 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.
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