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2016 ; 7
(5
): 1768-82
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Optimizing ultrafast illumination for multiphoton-excited fluorescence imaging
#MMPMID27231620
Stoltzfus CR
; Rebane A
Biomed Opt Express
2016[May]; 7
(5
): 1768-82
PMID27231620
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We study the optimal conditions for high throughput two-photon excited
fluorescence (2PEF) and three-photon excited fluorescence (3PEF) imaging using
femtosecond lasers. We derive relations that allow maximization of the rate of
imaging depending on the average power, pulse repetition rate, and noise
characteristics of the laser, as well as on the size and structure of the sample.
We perform our analysis using ~100 MHz, ~1 MHz and 1 kHz pulse rates and using
both a tightly-focused illumination beam with diffraction-limited image
resolution, as well loosely focused illumination with a relatively low image
resolution, where the latter utilizes separate illumination and fluorescence
detection beam paths. Our theoretical estimates agree with the experiments, which
makes our approach especially useful for optimizing high throughput imaging of
large samples with a field-of-view up to 10x10 cm(2).