Multi-dimensional fluorescence lifetime measurements

被引：0

作者：

Biskup, Christoph ^{[1
]}

Hoffmann, Birgit ^{[1
]}

Kelbauskas, Laimonas ^{[1
,2
]}

Zimmer, Thomas ^{[1
]}

Dietrich, Sascha ^{[1
]}

Becker, Wolfgang ^{[3
]}

Bergmann, Axel ^{[3
]}

Kloecker, Nikolaj ^{[4
]}

Benndorf, Klaus ^{[1
]}

机构：

[1] Univ Jena, Inst Phys 2, Teichgraben 8, D-07740 Jena, Germany

[2] Arizona State Univ, Biodesign Inst, Tempe, AZ 85287 USA

[3] Becker & Hickl GmbH, D-12277 Berlin, Germany

[4] Univ Freiburg, Inst Physiol 2, D-79104 Freiberg, Germany

来源：

MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES VII | 2007年 / 6442卷

关键词：

multi-wavelength fluorescence lifetime imaging (mwFLIM); Forster resonance energy transfer (FRET); time-correlated-single-photon-counting (TCSPC); streak camera;

D O I：

10.1117/12.699870

中图分类号：

R318 [生物医学工程];

学科分类号：

0831 ;

摘要：

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 single-photon-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 Forster 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.

引用

页数：12

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