Probing subtle fluorescence dynamics in cellular proteins by streak camera based fluorescence lifetime imaging microscopy

Krishnan, R. V.; Biener, Eva; Jian-Hua Zhang; Heckel, Robert; Herman, Brian
December 2003
Applied Physics Letters;12/1/2003, Vol. 83 Issue 22, p4658
Academic Journal
We report the cell biological applications of a recently developed multiphoton fluorescence lifetime imaging microscopy system using a streak camera (StreakFLIM). The system was calibrated with standard fluorophore specimens and was shown to have high accuracy and reproducibility. We demonstrate the applicability of this instrument in living cells for measuring the effects of protein targeting and point mutations in the protein sequence, which are not obtainable by conventional intensity-based fluorescence microscopy methods. We discuss the relevance of such time-resolved information in quantitative energy transfer microscopy and in measurement of the parameters that characterize the intracellular physiology. © 2003 American Institute of Physics.


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