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This paper was published in Optics Express and is made available as an electronic reprint with the permission of OSA. The paper can be found at the following URL on the OSA website: http://www.opticsinfobase.org/oe/abstract.cfm?uri=oe-18-13-13661. Systematic or multiple reproduction or distribution to multiple locations via electronic or other means is prohibited and is subject to penalties under law. See the OSA Author Copyright Permissions page for further information.

Abstract

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

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.

Citation:
Jeffrey J. Field, Ramón Carriles, Kraig E. Sheetz, Eric V. Chandler, Erich E. Hoover, Shane E. Tillo, Thom E. Hughes, Anne W. Sylvester, David Kleinfeld, and Jeff A. Squier, "Optimizing the fluorescent yield in two-photon laser scanning microscopy with dispersion compensation," Opt. Express 18, 13661-13672 (2010)

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

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