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Proceedings Paper

Frequency domain phosphorescence lifetime Imaging measurements and applications by ISS FastFLIM and multi pulse excitation
Author(s): Ulas C. Coskun; Sandra Lam; Yuansheng Sun; Shih-Chu Jeff Liao; Steven C. George; Beniamino Barbieri
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Paper Abstract

Phosphorescence probes can have significantly long lifetimes, on the order of micro- to milli-seconds or longer. In addition, environmental changes can affect the lifetimes of these phosphorescence probes. Thus, Phosphorescence Lifetime Imaging Microscopy (PLIM) is a very useful tool to localize the phosphorescence probes based on their lifetimes to study the variance in the lifetimes due to the micro environmental changes. Since the probes respond to the biologically relevant parameters like oxygen concentration, they can be used to study various biologically relevant processes like cellular metabolism, protein interaction etc. In this case, we study the effects of oxygen on Oxyphor G4 with PLIM. Since The Oxyphor G4 can be quenched by O2, it is a good example of such a probe and has a lifetime around 250us. Here we present the digital frequency domain PLIM technique and study the lifetime of the Oxyphor G4 as a function of the O2 concentration. The lifetime data are successfully presented in a phasor plot for various O2 concentrations and are consistent with the time domain data. Overall, we can analyze the oxygen consumption of varying cells using this technique.

Paper Details

Date Published: 21 February 2017
PDF: 8 pages
Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 1006918 (21 February 2017); doi: 10.1117/12.2255481
Show Author Affiliations
Ulas C. Coskun, ISS, Inc. (United States)
Sandra Lam, Washington Univ. in St. Louis (United States)
Yuansheng Sun, ISS, Inc. (United States)
Shih-Chu Jeff Liao, ISS, Inc. (United States)
Steven C. George, Washington Univ. in St. Louis (United States)
Beniamino Barbieri, ISS, Inc. (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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