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The « spectral » approach consists of analysing fluorescence variation by extracting quantitative
information from the entire fluorescence spectrum. It is ideally suited to technologies based on
fluorescence transfer such as FRET (Fluorescence Resonance Energy Transfer) in live cells.
Spectrum acquisition is particularly well adapted to early cellular signalling event analysis
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 high sample acquisition rates in the 50 ms time-range (ideal for short-time cellular events)
quantitative real-time protein translocation monitoring
cell imagery technique interpretation difficulties (morphological dependence etc.) avoided
statistically meaningful averaging over populations of 100s of cells
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discrimination between different biosensors
elimination of cell and culture plate auto-fluorescence, improving signal/noise ratio
photo-bleaching bias detection and correction
intrinsic sample fluorescence determination, eliminating false-positives
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Kinetic profiles : rapid spectra acquisition (ca. 20 points/sec) eliminates
false negatives associated with pathway signalling effects of short-term duration
Dose-response curves : accurate dose-response curves determined by curve
fitting of FRET time-course data
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