Page 83 - Vibrational Spectroscopic Imaging for Biomedical Applications

CHAPTER 3

Preparation of Tissues

and Cells for Infrared and

Raman Spectroscopy

and Imaging

Ehsan Gazi, Peter Gardner
Manchester Interdisciplinary Biocentre (MIB)
The University of Manchester
Manchester, United Kingdom

3.1 Introduction
Vibrational techniques, Raman and Fourier transform IR (FTIR)
microspectroscopy, provide structural information as well as relative
quantification of lipids, proteins, carbohydrates and a variety of
phosphorylated biomolecules within biological samples such as
whole mammalian cells or tissue. However, the full potential of these
technologies to interrogate this wide-range of biomolecules is only
realized if careful consideration is given to sample preparation. This
element of the experimental design can have significant implications
for the interpretation of spectra and thus for their biochemical rele-
vance as well as the spatial distribution of biomolecules in imaging
studies.
Cells are naturally present in hydrated form, whereby water
molecules are bound to macromolecules such as proteins, phospho-
lipids, and carbohydrates and this contributes to their structural
integrity and function. A review of the early literature concerning

59