Multiwavelength FLIM: new applications and algorithms

A. Rück; D. Strat; F. Dolp; B. von Einem; C. A. F. von Arnim

doi:10.1117/12.874648

28 February 2011 Multiwavelength FLIM: new applications and algorithms

A. Rück, D. Strat, F. Dolp, B. von Einem, C. A. F. von Arnim

Proceedings Volume 7903, Multiphoton Microscopy in the Biomedical Sciences XI; 790321 (2011) https://doi.org/10.1117/12.874648
Event: SPIE BiOS, 2011, San Francisco, California, United States

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.

Citation Download 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 (28 February 2011); https://doi.org/10.1117/12.874648

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