Measurement of the chirp-dependent fluorescence response: a window to excited-state population dynamics

G. J. Brakenhoff; Arjan H. Buist; Michiel Mueller; Jeffrey A. Squier; Chris J. Bardeen; Vladislav V. Yakovlev; Kent R. Wilson

doi:10.1117/12.347587

6 May 1999 Measurement of the chirp-dependent fluorescence response: a window to excited-state population dynamics

G. J. Brakenhoff, Arjan H. Buist, Michiel Mueller, Jeffrey A. Squier, Chris J. Bardeen, Vladislav V. Yakovlev, Kent R. Wilson

Author Affiliations +

Proceedings Volume 3605, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing VI; (1999) https://doi.org/10.1117/12.347587
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

Abstract

High intensity chirped pulses can be used for probing microscopic chemical environments through the use of a particular choice of dye, for instance SNAFL2. The basis for this technique is that the excited state populations can be manipulated through control over the temporal order of the excitation frequencies in the excitation pulse -- i.e. chirp - - with the outcoming fluorescence as the reporting parameter. A chirp dependent fluorescence response can also be observed in larger molecular systems with more degrees of freedom like for instance green fluorescent proteins. In preparation for application of the technique to microscopy we use a facility permitting observation of this phenomenon in various dyes with high sensitivity. High power, 30 fs pulses from an OPA, tunable from 400 nm to 1.5 micron are used. These pulses with a repetition rate of 1 kHz are sufficiently intense that a relatively large sample region can be excited to saturation from which then a sub-region with uniform excitation conditions can be selected for signal collection.

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G. J. Brakenhoff, Arjan H. Buist, Michiel Mueller, Jeffrey A. Squier, Chris J. Bardeen, Vladislav V. Yakovlev, and Kent R. Wilson "Measurement of the chirp-dependent fluorescence response: a window to excited-state population dynamics", Proc. SPIE 3605, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing VI, (6 May 1999); https://doi.org/10.1117/12.347587

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