10 February 2007 Multi-dimensional fluorescence lifetime measurements
Author Affiliations +
Proceedings Volume 6442, Multiphoton Microscopy in the Biomedical Sciences VII; 64420V (2007); doi: 10.1117/12.699870
Event: SPIE BiOS, 2007, San Jose, California, United States
In this study, we present two different approaches that allow multi-wavelength fluorescence lifetime measurements in the time domain. One technique is based on a streak camera system, the other technique is based on a time-correlated singlephoton- counting (TCSPC) approach. The setup consists of a confocal laser-scanning microscope (LSM 510, Zeiss) and a Titanium:Sapphire-laser (Mira 900D, Coherent) that is used for pulsed one- and two-photon excitation. Fluorescence light emitted by the sample is dispersed by a polychromator (250is, Chromex) and recorded by a streak camera (C5680 with M5677 sweep unit, Hamamatsu Photonics) or a 16 channel TCSPC detector head (PML-16, Becker & Hickl) connected to a TCSPC imaging module (SPC-730/SPC-830, Becker & Hickl). With these techniques it is possible to acquire fluorescence decays in several wavelength regions simultaneously. We applied these methods to Förster resonance energy transfer (FRET) measurements and discuss the advantages over fluorescence techniques that are already well established in the field of confocal microscopy, such as spectrally resolved intensity measurements or single-wavelength fluorescence lifetime measurements.
© (2007) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Christoph Biskup, Birgit Hoffmann, Laimonas Kelbauskas, Thomas Zimmer, Sascha Dietrich, Wolfgang Becker, Axel Bergmann, Nikolaj Klöcker, Klaus Benndorf, "Multi-dimensional fluorescence lifetime measurements", Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 64420V (10 February 2007); doi: 10.1117/12.699870; https://doi.org/10.1117/12.699870


Fluorescence resonance energy transfer




Streak cameras

Confocal microscopy


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