Multiphoton fluorescence microspectroscopy

Fu-Jen Kao; Bai-Ling Lin; Ping Chin Cheng

doi:10.1117/12.384177

2 May 2000 Multiphoton fluorescence microspectroscopy

Fu-Jen Kao, Bai-Ling Lin, Ping Chin Cheng

Proceedings Volume 3919, Three-Dimensional and Multidimensional Microscopy: Image Acquisition Processing VII; (2000) https://doi.org/10.1117/12.384177
Event: BiOS 2000 The International Symposium on Biomedical Optics, 2000, San Jose, CA, United States

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.

Citation Download Citation

Fu-Jen Kao, Bai-Ling Lin, and Ping Chin Cheng "Multiphoton fluorescence microspectroscopy", Proc. SPIE 3919, Three-Dimensional and Multidimensional Microscopy: Image Acquisition Processing VII, (2 May 2000); https://doi.org/10.1117/12.384177

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