Frequency domain phosphorescence lifetime Imaging measurements and applications by ISS FastFLIM and multi pulse excitation

Ulas C. Coskun, Sandra Lam, Yuansheng Sun, Shih Chu Jeff Liao, Steven George, Beniamino Barbieri

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Scopus citations


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.

Original languageEnglish (US)
Title of host publicationMultiphoton Microscopy in the Biomedical Sciences XVII
EditorsKarsten Konig, Peter T. C. So, Ammasi Periasamy, Xiaoliang S. Xie
ISBN (Electronic)9781510605794
StatePublished - Jan 1 2017
Externally publishedYes
EventMultiphoton Microscopy in the Biomedical Sciences XVII - San Francisco, United States
Duration: Jan 29 2017Jan 31 2017


OtherMultiphoton Microscopy in the Biomedical Sciences XVII
Country/TerritoryUnited States
CitySan Francisco

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Biomaterials
  • Radiology Nuclear Medicine and imaging


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