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Differential levels of metabolic activity in isolated versus confluent/partially confluent HeLa cells are analyzed by autofluorescent NAD(P)H using multi-photon FLIM microscopy
Author(s): Andrea Chandler; Aaron Chandler; Horst Wallrabe; Ammasi Periasamy
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Paper Abstract

NAD(P)H is a known biomarker for cellular metabolism; a higher ratio of enzyme-bound NAD(P)H to free/unbound NAD(P)H indicates an increase in metabolic activity. Free NADH has a shorter fluorescence lifetime (τ1), the bound version (τ2) a longer lifetime. FLIM’s unique capability to establish inter alia the relative fractions of τ1 (a1%) and τ2 (a2%) in each pixel, determines the level of metabolic activity. The relative abundances of bound NAD(P)H were analyzed for single cells, confluent and partially confluent cells within 3 Fields-of-View (FoVs). A gradient of increasing a 2% levels of bound NAD(P)H from single, partially confluent to confluent cells was observed.

Paper Details

Date Published: 2 March 2017
PDF: 6 pages
Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 100692R (2 March 2017); doi: 10.1117/12.2267657
Show Author Affiliations
Andrea Chandler, Palo Alto High School (United States)
Aaron Chandler, Univ. of California, Davis (United States)
Horst Wallrabe, Univ. of Virginia (United States)
Ammasi Periasamy, Univ. of Virginia (United States)


Published in SPIE Proceedings Vol. 10069:
Multiphoton Microscopy in the Biomedical Sciences XVII
Ammasi Periasamy; Peter T. C. So; Karsten König; Xiaoliang S. Xie, Editor(s)

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