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Opt Express. 2010 Jun 21;18(13):13661-72. doi: 10.1364/OE.18.013661.

Optimizing the fluorescent yield in two-photon laser scanning microscopy with dispersion compensation.

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Center for Microintegrated Optics for Advanced Bioimaging and Control, and Department of Physics, Colorado School of Mines, Golden, CO 80401, USA.


A challenge for nonlinear imaging in living tissue is to maximize the total fluorescent yield from each fluorophore. We investigated the emission rates of three fluorophores-rhodamine B, a red fluorescent protein, and CdSe quantum dots-while manipulating the phase of the laser excitation pulse at the focus. In all cases a transform-limited pulse maximized the total yield to insure the highest signal-to-noise ratio. Further, we find evidence of fluorescence antibleaching in quantum dot samples.

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