Förster resonance energy transfer (FRET)-based picosecond lifetime reference for instrument response evaluation

R. Luchowski, P. Kapusta, M. Szabelski, P. Sarkar, Julian Borejdo, Zygmunt Gryczynski, Ignacy Gryczynski

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Förster resonance energy transfer (FRET) can be utilized to achieve ultrashort fluorescence responses in time-domain fluorometry. In a poly(vinyl) alcohol matrix, the presence of 60 mM Rhodamine 800 acceptor shortens the fluorescence lifetime of a pyridine 1 donor to about 20 ps. Such a fast fluorescence response is very similar to the instrument response function (IRF) obtained using scattered excitation light. A solid fluorescent sample (e.g a film) with picosecond lifetime is ideal for IRF measurements and particularly useful for time-resolved microscopy. Avalanche photodiode detectors, commonly used in this field, feature color- dependent-timing responses. We demonstrate that recording the fluorescence decay of the proposed FRET-based reference sample yields a better IRF approximation than the conventional light-scattering method and therefore avoids systematic errors in decay curve analysis.

Original languageEnglish
Article number095601
JournalMeasurement Science and Technology
Volume20
Issue number9
DOIs
StatePublished - 1 Jan 2009

Keywords

  • FRET
  • Impulse response function
  • Lifetime standard

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