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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.
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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