Effect of handling and fixation processes on fluorescence spectroscopy of mouse skeletal muscles under two-photon
M G Xu1, E D Williams, E W Thompson
1Centre for Micro-Photonics, School of Biophysical Sciences and Electrical Engineering, Swinburne University of Technology, P.O. Box 218 Hawthorn, 3122 Victoria, Australia.
Abstract:
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.
