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Applied Optics

Applied Optics


  • Vol. 39, Iss. 34 — Dec. 1, 2000
  • pp: 6312–6317

Effect of handling and fixation processes on fluorescence spectroscopy of mouse skeletal muscles under two-photon excitation

Ming G. Xu, Elizabeth D. Williams, Erik W. Thompson, and Min Gu  »View Author Affiliations

Applied Optics, Vol. 39, Issue 34, pp. 6312-6317 (2000)

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We investigated the effects of handling and fixation processes on the two-photon fluorescence spectroscopy of endogenous fluorophors in mouse skeletal muscle. The skeletal muscle was handled in one of two ways: either sectioned without storage or sectioned following storage in a freezer. The two-photon fluorescence spectra measured for different storage or fixation periods show a differential among those samples that were stored in water or were fixed either in formalin or methanol. The spectroscopic results indicate that formalin was the least disruptive fixative, having only a weak effect on the two-photon fluorescence spectroscopy of muscle tissue, whereas methanol had a significant influence on one of the autofluorescence peaks. The two handling processes yielded similar spectral information, indicating no different effects between them.

© 2000 Optical Society of America

OCIS Codes
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.4580) Medical optics and biotechnology : Optical diagnostics for medicine
(170.6280) Medical optics and biotechnology : Spectroscopy, fluorescence and luminescence
(170.6510) Medical optics and biotechnology : Spectroscopy, tissue diagnostics
(170.6930) Medical optics and biotechnology : Tissue
(170.7160) Medical optics and biotechnology : Ultrafast technology

Original Manuscript: March 29, 2000
Revised Manuscript: July 7, 2000
Published: December 1, 2000

Ming G. Xu, Elizabeth D. Williams, Erik W. Thompson, and Min Gu, "Effect of handling and fixation processes on fluorescence spectroscopy of mouse skeletal muscles under two-photon excitation," Appl. Opt. 39, 6312-6317 (2000)

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