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Proceedings Paper

Combined spectrally-resolved multiphoton microscopy and transmission microscopy employing a high-sensitivity electron-multiplying CCD camera
Author(s): Valeric? Raicu; Russell Fung; Mike Melnichuk; Anurag Chaturvedi; Devin Gillman
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Paper Abstract

We have developed a fast, sensitive multiphoton microscope employing a tunable femtosecond laser and an electron-multiplying CCD camera (EMCCD) in a non-descanned detection scheme. In one configuration, the microscope provides "standard" multiphoton fluorescence images with a relatively high acquisition rate (limited only by the speed of the scanners) over a broad band of emission wavelengths. In a second configuration, the use of a diffractive optic element allows acquisition of one full set of spectrally-resolved images after only one complete scan. Increased speed is achieved in this configuration by setting the integration time of the camera such that an entire line is scanned at once, with the spectrum at each point being perpendicular to the scanned line. In both configurations, a suitable optical arrangement permits acquisition of high resolution images for both multiphoton excitation of fluorescence and widefield transmission microscopy. To illustrate the performances of our microscope we present spectrally-resolved images of individual fluorescent molecules spread over a surface as well as fluorescence images of yeast cells expressing a membrane receptor tagged with a variant of the green fluorescent protein.

Paper Details

Date Published: 10 February 2007
PDF: 5 pages
Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 64420M (10 February 2007); doi: 10.1117/12.705920
Show Author Affiliations
Valeric? Raicu, Univ. of Wisconsin, Milwaukee (United States)
Russell Fung, Univ. of Wisconsin, Milwaukee (United States)
Mike Melnichuk, Univ. of Wisconsin, Milwaukee (United States)
Anurag Chaturvedi, Univ. of Wisconsin, Milwaukee (United States)
Devin Gillman, Univ. of Wisconsin, Milwaukee (United States)

Published in SPIE Proceedings Vol. 6442:
Multiphoton Microscopy in the Biomedical Sciences VII
Ammasi Periasamy; Peter T. C. So, Editor(s)

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