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Fluorescence spectroscopy for revealing mechanisms in biology: Strengths and pitfalls

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Abstract

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.

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Figure 1

Reproduced with permission from Swaminathan et al. (1996).

Figure 2

Figure reproduced with permission from Lakshmikanth et al. (2001).

Figure 3

Reproduced with permission from Bhatia et al. (2018).

Figure 4

Reproduced with permission from Bhatia et al. (2018).

Figure 5
Figure 6

Reproduced with permission from Biswas et al. (2017).

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Acknowledgements

Collaboration with Prof Jayant Udgaonkar, Prof Ruchi Anand, Prof R Swaminathan, Dr G Lakshmikanth, Dr K Sridevi, Dr Tapan Das, Dr Arvind Srivastava, Ms Sandhya Bhatia, Dr Anwesha Biswas and Dr Jessy Miriam John is gratefully acknowledged. Their work is described in some detail here. I thank Prof N Periasamy for providing the software used in the analysis of data shown here.

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Authors and Affiliations

  1. Department of Biotechnology, Anna University, Chennai, 600 025, India

    G Krishnamoorthy

  2. Department of Chemistry, Indian Institute of Technology, Mumbai, 400 076, India

    G Krishnamoorthy

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  1. G Krishnamoorthy
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Correspondence to G Krishnamoorthy.

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Krishnamoorthy, G. Fluorescence spectroscopy for revealing mechanisms in biology: Strengths and pitfalls. J Biosci 43, 555–567 (2018). https://doi.org/10.1007/s12038-018-9763-4

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