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Proceedings Paper

Monitoring changes in endogenous fluorophores through quantitative FLIM imaging in live cells
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Paper Abstract

Changes in energy metabolism, mitochondrial functions and of reactive oxygen species are often supposed to induce alterations in cellular activity. The major intracellular endogenous fluorophores are reduced nicotinamide adenine dinucleotide (NADH) and dinucleotide phosphate (NADPH), riboflavin's, and tryptophan present inside biological tissue and they can be used to image tissue architecture without any exogenous probe. Their fluorescence can be excited by multi-photon microscopy using NIR laser wavelengths [1,2,5,6]. Using FLIM imaging the lifetime of tryptophan and NADH were monitored in cells with and without addition of glucose in the medium. The lifetime data were collected and further using the ANOVA (refer table 2) of the lifetime of free NADH, bound NADH and Tryptophan, we found on applying the null hypothesis for the P-value > 0.05, there is a significant difference between the lifetimes of bound NADH and Tryptophan from control to glucose treated cells, however, free NADH, shown to be not significant change between the control and glucose treated cells. The tryptophan puzzle comes closer and closer to a solution. The ultimate evidence supporting the existence of FRET between Tryptophan and NADH in live cells slowly could come from lifetime measurements of tryptophan in proteins and bound NADH within live cells.

Paper Details

Date Published: 9 February 2012
PDF: 7 pages
Proc. SPIE 8226, Multiphoton Microscopy in the Biomedical Sciences XII, 822644 (9 February 2012); doi: 10.1117/12.923666
Show Author Affiliations
Vinod Jyothikumar, Univ. of Virginia (United States)
Yuansheng Sun, Univ. of Virginia (United States)
Ammasi Periasamy, Univ. of Virginia (United States)

Published in SPIE Proceedings Vol. 8226:
Multiphoton Microscopy in the Biomedical Sciences XII
Karsten König, Editor(s)

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