Tear Down the Fluorescent Curtain: A New Fluorescence Suppression Method for Raman Microspectroscopic Analyses

The near exponential proliferation of published Raman microspectroscopic applications over the last decade bears witness to the strengths and versatility of this technology. However, laser-induced fluorescence often severely impedes its application to biological samples. Here we report a new approac...

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Bibliographic Details
Published in:Scientific reports 2019-10, Vol.9 (1), p.15785-9, Article 15785
Main Authors: Yakubovskaya, Elena, Zaliznyak, Tatiana, Martínez Martínez, Joaquin, Taylor, Gordon T.
Format: Article
Language:English
Subjects:
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Algae
Aquatic microorganisms
Biological samples
Carotenoids
Fluorescence
Humanities and Social Sciences
Hydrogen peroxide
Irradiation
Isotopic tracers
Lasers
Macromolecules
Methods
multidisciplinary
Oxidation
Questioning
Radiation
Science
Science (multidisciplinary)
Spectrum analysis
Tracers
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Summary:The near exponential proliferation of published Raman microspectroscopic applications over the last decade bears witness to the strengths and versatility of this technology. However, laser-induced fluorescence often severely impedes its application to biological samples. Here we report a new approach for near complete elimination of laser-induced background fluorescence in highly pigmented biological specimens (e.g., microalgae) enabling interrogation by Raman microspectroscopy. Our simple chemiphotobleaching method combines mild hydrogen peroxide oxidation with broad spectrum visible light irradiation of the entire specimen. This treatment permits observing intracellular distributions of macromolecular pools, isotopic tracers, and even viral propagation within cells previously not amenable to Raman microspectroscopic examination. Our approach demonstrates the potential for confocal Raman microspectroscopy becoming an indispensable tool to obtain spatially-resolved data on the chemical composition of highly fluorescent biological samples from individual cells to environmental samples.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-019-52321-3