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Proceedings Paper

Fluorescence lifetime imaging (FLIM): simulation and error analysis
Author(s): Masilamani Elangovan; Peter Kionga-Kamau; Ammasi Periasamy
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Paper Abstract

Real-time monitoring of reorganization and molecular interactions of live cellular components on a precise spatial and temporal scale, requires a high-speed and high- sensitivity imaging system. Theoretically, excited state fluorescence lifetime imaging techniques provide high temporal resolution for dynamics studies of biological samples. In order to provide a comprehensive foundation for the development of the technique, we simulated images using the decay and normal equations for a single component with different windowing schemes. In this model we varied several parameters involved in the simulation to produce images under different theoretical conditions. To obtain a realistic result, we considered equations for noise due to readout, dark current, photon shot noises, and other factors. In addition, the simulant was validated using sample decay images with known lifetimes, which were obtained with current fluorescent decay systems. The software developed through this research is intended to create a compete tutorial describing the theory and procedures of lifetime image acquisition. This theoretical simulation is verified with experiments using known fluorophore lifetimes.

Paper Details

Date Published: 27 April 2000
PDF: 8 pages
Proc. SPIE 3921, Optical Diagnostics of Living Cells III, (27 April 2000); doi: 10.1117/12.384230
Show Author Affiliations
Masilamani Elangovan, Univ. of Virginia (United States)
Peter Kionga-Kamau, Univ. of Virginia (United States)
Ammasi Periasamy, Univ. of Virginia (United States)

Published in SPIE Proceedings Vol. 3921:
Optical Diagnostics of Living Cells III
Daniel L. Farkas; Daniel L. Farkas; Robert C. Leif, Editor(s)

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