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Multi-Photon Fluorescence Spectroum of Common Nucleic Acid Probes

Published online by Cambridge University Press:  02 July 2020

P. C. Cheng ,
B. L. Lin ,
F. J. Kao ,
C. K. Sun  and
I. Johnson
P. C. Cheng
Affiliation:
AMIL, Dept. of Electrical Engineering, Univ. at Buffalo, Buffalo, NY14260USA
B. L. Lin
Affiliation:
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, 11529Republic of China
F. J. Kao
Affiliation:
Department of Physics, National Sun Yat-sen University, Kaohsiung, Taiwan, 80424Republic of China
C. K. Sun
Affiliation:
Department of Electrical Engineering, National Taiwan University, Taipei, Taiwan, 10609Rep. of China
I. Johnson
Affiliation:
Molecular Probes Inc., Eugene, OR , 97402. USA
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Extract

Fluorescent probes are commonly used in biological fluorescence microscopy for tracking specific structures and sub-cellular compartments, and for indicating cellular ionic conditions. Recent development in multi-photon fluorescence microscopy has greatly expanded the usage of fluorescent probes in biomedical research. Considering its non-linear nature, two-photon excitation may generate very different fluorescence spectral response in the sample when compared with single photon excitation. It is thus necessary to measure the two-photon spectra of various fluorescent probes, so that two-photon fluorescence microscopy may be operated effectively and the images properly interpreted. This report represents the first installment of a continued effort in characterizing the multi-photon fluorescence spectra of commonly used bio-probes.

Two-photon fluorescence spectra excited with near infrared at 780nm were obtained with a SpectraPro-500 spectrophotometer (Acton Research) equipped with a TE-cooled PMT and coupled to a Spectra-Physics Tsunami Ti-sapphire laser pumped by a Coherent Verdi solid-state laser operated at 85MHz, l00fs pulse.

Type
Confocal Microscopy
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 820 - 821
Copyright
Copyright © Microscopy Society of America

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References

References:

1.Cheng, P. C., et al., J. Microscopy, 189, 199–212. (1998).CrossRefGoogle Scholar
2.Kao, F. J. et al., SPIE Proceedings, 3919 (2000) in pressGoogle Scholar
3.Kao, F. J. et al., this volumeGoogle Scholar
This project was support by Academia Sinica (BLL), the National Science Council, Rep. of China [NSC-88-2811-B-001-0023 (PCC), NSC-89-2311-B-001-032 (BLL), NSC89-2112-M-l 10-016, NSC89-2216-E-110-003 (FJK)] and Mr. and Mrs. Jin-Mu Huang of Aurum Belle Investment Co. (on behalf of the Ge-An Charity, ) Fluorescence stains used in this study were provided by Molecular Probes, Inc. Eugene, OR, USA.Google Scholar