Multidimensional fluorescence lifetime measurements

Christoph Biskup; Birgit Hoffmann; Laimonas Kelbauskas; Thomas Zimmer; Nikolaj Klöcker; Wolfgang Becker; Klaus Benndorf

doi:10.1117/12.762834

15 February 2008 Multidimensional fluorescence lifetime measurements

Christoph Biskup, Birgit Hoffmann, Laimonas Kelbauskas, Thomas Zimmer, Nikolaj Klöcker, Wolfgang Becker, Klaus Benndorf

Proceedings Volume 6860, Multiphoton Microscopy in the Biomedical Sciences VIII; 68601I (2008) https://doi.org/10.1117/12.762834
Event: SPIE BiOS, 2008, San Jose, California, United States

Abstract

In this study, we present two different approaches that can be used for multi-wavelength fluorescence lifetime measurements in the time domain. One technique is based on a streak-camera system, the other technique is based on the time-correlated-single-photon-counting (TCSPC) approach. The setup consists of a confocal laser-scanning microscope and a Titanium:Sapphire-laser that is used for pulsed one- and two-photon excitation. Fluorescence light emitted by the sample is fed back through the scan head and guided to one of the confocal channels, where it is coupled into an optical fiber and directed to a polychromator. The polychromator disperses the emitted light according to its wavelength and focuses the resulting spectrum on the entrance slit of a streak camera or a 16 channel PMT array, which is connected to a TCSPC imaging module. 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 and pitfalls of fluorescence lifetime measurements.

Citation Download Citation

Christoph Biskup, Birgit Hoffmann, Laimonas Kelbauskas, Thomas Zimmer, Nikolaj Klöcker, Wolfgang Becker, and Klaus Benndorf "Multidimensional fluorescence lifetime measurements", Proc. SPIE 6860, Multiphoton Microscopy in the Biomedical Sciences VIII, 68601I (15 February 2008); https://doi.org/10.1117/12.762834

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

Proteins

Molecules

Microscopes

Confocal microscopy

Streak cameras

Show All Keywords

Keywords/Phrases

Search In:

Publication Years