Fluorescence Lifetime Correlation Spectroscopy

Fluorescence lifetime correlation spectroscopy ( English fluorescence lifetime correlation spectroscopy , FLCS) combines the time-correlated single photon counting (engl. Time-correlated single photon counting , TCSPC) with the classical fluorescence correlation spectroscopy (engl. Fluorescence Correlation Spectroscopy , FCS).

With this method, the time-resolved ( picosecond range) detection of the fluorescence is used to separate the contribution of different processes to the measurement signal (e.g. different lifetimes of different dyes or separation of afterpulsing or dark noise of the detector from the actual measurement signal). The method is based on the assumption that the different emission processes have different decay times and thus different TCSPC histograms , whereby the individual intensity components can be statistically separated from one another by weighting.

In contrast to classic FCS, the prerequisite for the FLCS is pulsed excitation (pulses in the picosecond range) and the simultaneous measurement of the arrival times of the photons on two time scales (with picosecond accuracy relative to the excitation pulse and with nanosecond accuracy relative to the start of the measurement ). This is used, for example, in biophysics to determine protein-protein interactions .

literature

Martin Böhmer, Michael Wahl, Hans-Jürgen Rahn, Rainer Erdmann, Jörg Enderlein: Time-resolved fluorescence correlation spectroscopy . In: Chemical Physics Letters . tape 353 , no. 5-6 , February 26, 2002, pp. 439-445 , doi : 10.1016 / S0009-2614 (02) 00044-1 .
Jörg Enderlein, Ingo Gregor: Using fluorescence lifetime for discriminating detector after pulsing in fluorescence-correlation spectroscopy . In: Review of Scientific Instruments . tape 76 , no. 3 , February 15, 2005, p. 033102 , doi : 10.1063 / 1.1863399 .
Peter Kapusta, Michael Wahl, Aleš Benda, Martin Hof, Jörg Enderlein: Fluorescence Lifetime Correlation Spectroscopy . In: Journal of Fluorescence . tape 17 , no. 1 , December 14, 2006, p. 43-48 , doi : 10.1007 / s10895-006-0145-1 .

Web links

Overview of fluorescence lifetime correlation spectroscopy from PicoQuant GmbH (English)

Individual evidence

^ Y. Tian, MM Martinez, D. Pappas: Fluorescence correlation spectroscopy: a review of biochemical and microfluidic applications. In: Applied spectroscopy. Volume 65, Number 4, April 2011, ISSN 1943-3530 , pp. 115A-124A, doi: 10.1366 / 10-06224 , PMID 21396180 , PMC 3071976 (free full text).
^ RN Day, F. Schaufele: Imaging molecular interactions in living cells. In: Molecular endocrinology (Baltimore, Md.). Volume 19, Number 7, July 2005, ISSN 0888-8809 , pp. 1675-1686, doi: 10.1210 / me.2005-0028 , PMID 15761028 . PMC 2900770 (free full text).

[1] Y. Tian, MM Martinez, D. Pappas: Fluorescence correlation spectroscopy: a review of biochemical and microfluidic applications. In: Applied spectroscopy. Volume 65, Number 4, April 2011, ISSN 1943-3530 , pp. 115A-124A, doi: 10.1366 / 10-06224 , PMID 21396180 , PMC 3071976 (free full text).

[2] RN Day, F. Schaufele: Imaging molecular interactions in living cells. In: Molecular endocrinology (Baltimore, Md.). Volume 19, Number 7, July 2005, ISSN 0888-8809 , pp. 1675-1686, doi: 10.1210 / me.2005-0028 , PMID 15761028 . PMC 2900770 (free full text).