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Proceedings Article

Fast algorithms for the analysis of spectral FLIM data

[+] Author Affiliations
I. Gregor, J. Enderlein

Georg-August-Univ. (Germany)

B. Krämer, F. Koberling, R. Erdmann, M. Wahl

PicoQuant GmbH (Germany)

S. Fore

PicoQuant Photonics North America, Inc. (USA)

Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 790330 (February 10, 2011); doi:10.1117/12.873386
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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 simultaneous spectral detection together with Fluorescence Lifetime Imaging (sFLIM) allows collecting the complete information inherent to the fluorescence signal. Their fingerprint of lifetime and spectral properties identify the fluorescent labels unambiguously. Multiple labels can be investigated in parallel and separated from inherent auto-fluorescence of the sample. In addition, spectral FLIM FRET has the prospect to allow simultaneous detection of multiple FRET signals with quantitative analysis of FRET-efficiency and degree of binding. Spectral FLIM measurements generate huge amount of data. Suitable analysis procedures must be found to condense the inherent information to answer the scientific questions in a straightforward way. Different analysis techniques have been evaluated for a diversity of applications as multiplex labeling, quantitative determination of environmental parameters and distance measurements via FLIM FRET. In order to reach highest sensitivity in single photon detection, different detector types are investigated and developed. SPAD arrays equipped with micro-lenses promise superior detection efficiency while the integration of a spectrograph with a PMT array is easier to realize and allows for a higher number of detection channels. High detection speed can be realized through parallel TCSPC channels. In order to overcome the limits of the USB 2.0 interface, new interface solutions have been realized for the multichannel TCSPC unit HydraHarp 400.

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

I. Gregor ; B. Krämer ; F. Koberling ; R. Erdmann ; J. Enderlein, et al.
"Fast algorithms for the analysis of spectral FLIM data", Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 790330 (February 10, 2011); doi:10.1117/12.873386; http://dx.doi.org/10.1117/12.873386


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