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Proceedings Paper

Multiphoton fluorescence microspectroscopy
Author(s): Fu-Jen Kao; Bai-Ling Lin; Ping Chin Cheng
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Paper Abstract

The intrinsic confined photo-interacting volume in multi- photon fluorescence microscopy provides the possibility of obtaining fluorescence spectrum from specific cellular structure in a tissue. In this article, we demonstrated that it is feasible to obtain useful two-photon pumped fluorescence spectrum from cell wall and single chloroplast. The difference in fluorescence spectra obtained with single- and two-photon excitation indicates that a significant shift in fluorescence maximum may occur due to the non-linear nature of excitation. Therefore, in order to properly interpret two-photon fluorescence micrographs, it is important to characterize the fluorescence spectrum of the specimen and the commonly used fluorescence probes. The fluorescence spectra will in turn be useful in the selection of filter sets in multi-photon fluorescence microscopy.

Paper Details

Date Published: 2 May 2000
PDF: 7 pages
Proc. SPIE 3919, Three-Dimensional and Multidimensional Microscopy: Image Acquisition Processing VII, (2 May 2000); doi: 10.1117/12.384177
Show Author Affiliations
Fu-Jen Kao, National Sun Yat-sen Univ. (Taiwan)
Bai-Ling Lin, Institute of Molecular Biology (Taiwan)
Ping Chin Cheng, SUNY/Buffalo (United States)


Published in SPIE Proceedings Vol. 3919:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition Processing VII
Jose-Angel Conchello; Carol J. Cogswell; Andrew G. Tescher; Tony Wilson, Editor(s)

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