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Quantitative comparison between full‐spectrum and filter‐based imaging in hyperspectral fluorescence microscopy
Author(s) -
GAO L.,
HAGEN N.,
TKACZYK T.S.
Publication year - 2012
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2012.03596.x
Subject(s) - hyperspectral imaging , spectral imaging , microscope , full spectral imaging , microscopy , filter (signal processing) , optics , chemical imaging , context (archaeology) , signal (programming language) , materials science , fluorescence microscope , imaging spectroscopy , optical filter , dynamic range , fluorescence , computer science , computer vision , physics , artificial intelligence , paleontology , biology , programming language
Summary We implement a filterless illumination scheme on a hyperspectral fluorescence microscope to achieve full‐range spectral imaging. The microscope employs polarisation filtering, spatial filtering and spectral unmixing filtering to replace the role of traditional filters. Quantitative comparisons between full‐spectrum and filter‐based microscopy are provided in the context of signal dynamic range and accuracy of measured fluorophores’ emission spectra. To show potential applications, a five‐colour cell immunofluorescence imaging experiment is theoretically simulated. Simulation results indicate that the use of proposed full‐spectrum imaging technique may result in three times improvement in signal dynamic range compared to that can be achieved in the filter‐based imaging.