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Proceedings Article

Multiwavelength FLIM: new applications and algorithms

[+] Author Affiliations
A. Rück, D. Strat, F. Dolp

Institute for Laser Technologies in Medicine and Metrology (Germany)

B. von Einem, C. A. F. von Arnim

Univ. Ulm (Germany)

Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 790321 (February 28, 2011); doi:10.1117/12.874648
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From Conference Volume 7903

  • Multiphoton Microscopy in the Biomedical Sciences XI
  • Ammasi Periasamy; Karsten König; Peter T. C. So
  • San Francisco, California, USA | January 22, 2011

abstract

The combination of time-resolved and spectral resolved techniques as achieved by SLIM (spectrally resolved fluorescence lifetime imaging) improves the analysis of complex situations, when different fluorophores have to be distinguished. This could be the case when endogenous fluorophores of living cells and tissues are observed to identify the redox state and oxidative metabolic changes of the mitochondria. Other examples are FRET (resonant energy transfer) measurements, when different donor/acceptor pairs are observed simultaneously. SLIM is working in the time domain employing excitation with short light pulses and detection of the fluorescence intensity decay in many cases with time-correlated single photon counting (TCSPC). Spectral resolved detection is achieved by a polychromator in the detection path and a 16-channel multianode photomultiplier tube with the appropriate routing electronics. Within this paper special attention will be focused on FRET measurements with respect to protein interactions in Alzheimers disease. Using global analysis as the phasor plot approach or integration of the kinetic equations taking into account the multidimensional datasets in every spectral channel we could demonstrate considerable improvement of our calculations.

© (2011) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation

A. Rück ; D. Strat ; F. Dolp ; B. von Einem and C. A. F. von Arnim
"Multiwavelength FLIM: new applications and algorithms", Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 790321 (February 28, 2011); doi:10.1117/12.874648; http://dx.doi.org/10.1117/12.874648


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