Fluorescence spectroscopy for revealing mechanisms in biology: Strengths and pitfalls

This article describes the basic principles of steady-state and time-resolved fluorescence. The formal equivalence of the two methodologies is described first, followed by the extra advantages of time-resolved methods in revealing population heterogeneity in complex systems encountered in biology. S...

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Bibliographic Details
Published in:Journal of biosciences 2018-07, Vol.43 (3), p.555-567
Main Author: Krishnamoorthy, G
Format: Article
Language:English
Subjects:
Analytical methods
Bacillus amyloliquefaciens - chemistry
Bacillus amyloliquefaciens - ultrastructure
Bacterial Proteins - chemistry
Bacterial Proteins - metabolism
Benzopyrans - chemistry
Biology
Biomedical and Life Sciences
Biomedicine
Cell Biology
Complex systems
DNA-Binding Proteins - chemistry
DNA-Binding Proteins - metabolism
Fluorescence
Fluorescence Polarization
Fluorescence Recovery After Photobleaching
Fluorescence spectroscopy
Fluorescent Dyes - chemistry
Heterogeneity
Hydrogen-Ion Concentration
Life Sciences
Membrane Potentials - physiology
Microbiology
Naphthols - chemistry
Plant Proteins - chemistry
Plant Proteins - metabolism
Plant Sciences
Ranunculaceae - chemistry
Ranunculaceae - ultrastructure
Rhodamines - chemistry
Saccharomyces cerevisiae - chemistry
Saccharomyces cerevisiae - ultrastructure
Saccharomyces cerevisiae Proteins - chemistry
Saccharomyces cerevisiae Proteins - metabolism
Spectrometry, Fluorescence - methods
Tryptophan - chemistry
Tryptophan - metabolism
Zoology
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Description
Summary:This article describes the basic principles of steady-state and time-resolved fluorescence. The formal equivalence of the two methodologies is described first, followed by the extra advantages of time-resolved methods in revealing population heterogeneity in complex systems encountered in biology. Several examples from the author’s work are described in support of the above contention. Finally, several misinterpretations and pitfalls in the interpretation of fluorescence data and their remedy are described.
ISSN:0250-5991
0973-7138
DOI:10.1007/s12038-018-9763-4