Abstract
Fourier Transform Infrared (FTIR) micro-spectroscopy is an emerging technique for the biochemical analysis of tissues and cellular materials. It provides objective information on the holistic biochemistry of a cell or tissue sample and has been applied in many areas of medical research. However, it has become apparent that how the tissue is handled prior to FTIR micro-spectroscopic imaging requires special consideration, particularly with regards to methods for preservation of the samples. We have performed FTIR micro-spectroscopy on rodent heart and liver tissue sections (two spectroscopically very different biological tissues) that were prepared by desiccation drying, ethanol substitution and formalin fixation and have compared the resulting spectra with that of fully hydrated freshly excised tissues. We have systematically examined the spectra for any biochemical changes to the native state of the tissue caused by the three methods of preparation and have detected changes in infrared (IR) absorption band intensities and peak positions. In particular, the position and profile of the amide I, key in assigning protein secondary structure, changes depending on preparation method and the lipid absorptions lose intensity drastically when these tissues are hydrated with ethanol. Indeed, we demonstrate that preserving samples through desiccation drying, ethanol substitution or formalin fixation significantly alters the biochemical information detected using spectroscopic methods when compared to spectra of fresh hydrated tissue. It is therefore imperative to consider tissue preparative effects when preparing, measuring, and analyzing samples using FTIR spectroscopy.
Highlights
Fourier transform infrared (FTIR) spectroscopy and imaging is an emerging technique in the field of ex vivo diagnostics
In the last twenty years, the infrared spectra of single cells and intact tissues originating from dozens of species and cell types have been analyzed by several groups
It has become apparent that how the tissue is handled prior to FTIR imaging requires a number of considerations that are relatively unimportant when using conventional microscopy methods [22,23,24]
Summary
Fourier transform infrared (FTIR) spectroscopy and imaging is an emerging technique in the field of ex vivo diagnostics. In the last twenty years, the infrared spectra of single cells and intact tissues originating from dozens of species and cell types have been analyzed by several groups. Tissue Preservation Methods in Spectroscopy no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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