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Imaging spectrometer fundamentals for researchers in the biosciences—A tutorial
Author(s) -
Lerner Jeremy M.
Publication year - 2006
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20242
Subject(s) - spectrometer , spectral imaging , spectroscopy , optics , laser , confocal , spectral resolution , computer science , physics , fluorescence , fluorescence spectroscopy , spectral line , quantum mechanics , astronomy
Over the last 2 years there has been a dramatic increase in the number of bioscience laboratories using wavelength dispersive spectroscopy to study in vivo, in situ fluorescence. Transforming spectral information into an image provides a graphic means of mapping localized ionic, molecular, and protein–protein interactions. Spectroscopy also enables fluorophores with overlapping spectral features to be delineation. In this study, we provide the tools that a researcher needs to put into perspective instrumental contributions to a reported spectrum in order to gain greater understanding of the natural emission of the sample. We also show how to deduce the basic capabilities of a spectral confocal system. Finally, we show how to determine the true spectral bandwidth of an object, the illuminated area of a laser‐excited object, and what is needed to optimize light throughput. © 2006 International Society for Analytical Cytology