Multidimensional fluorescence lifetime measurements

Christoph Biskup; Birgit Hoffmann; Laimonas Kelbauskas; Thomas Zimmer; Sascha Dietrich; Wolfgang Becker; Axel Bergmann; Nikolaj Klöcker; Klaus Benndorf

doi:10.1117/12.699870

10 February 2007 Multi-dimensional fluorescence lifetime measurements

Christoph Biskup, Birgit Hoffmann, Laimonas Kelbauskas, Thomas Zimmer, Sascha Dietrich, Wolfgang Becker, Axel Bergmann, Nikolaj Klöcker, Klaus Benndorf

Author Affiliations +

Proceedings Volume 6442, Multiphoton Microscopy in the Biomedical Sciences VII; 64420V (2007) https://doi.org/10.1117/12.699870
Event: SPIE BiOS, 2007, San Jose, California, United States

Abstract

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.

Citation Download Citation

Christoph Biskup, Birgit Hoffmann, Laimonas Kelbauskas, Thomas Zimmer, Sascha Dietrich, Wolfgang Becker, Axel Bergmann, Nikolaj Klöcker, and Klaus Benndorf "Multi-dimensional fluorescence lifetime measurements", Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 64420V (10 February 2007); https://doi.org/10.1117/12.699870

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
12 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Luminescence

Fluorescence resonance energy transfer

Molecules

Proteins

Microscopes

Streak cameras

Confocal microscopy

Show All Keywords

Keywords/Phrases

Search In:

Publication Years