Chapter 10 Infrared and Raman Microscopy in Cell Biology

Abstract

This chapter presents novel microscopic methods to monitor cell biological processes of live or fixed cells without the use of any dye, stains, or other contrast agent. These methods are based on spectral techniques that detect inherent spectroscopic properties of biochemical constituents of cells, or parts thereof. Two different modalities have been developed for this task. One of them is infrared micro-spectroscopy, in which an average snapshot of a cell's biochemical composition is collected at a spatial resolution of typically 25 mm. This technique, which is extremely sensitive and can collect such a snapshot in fractions of a second, is particularly suited for studying gross biochemical changes. The other technique, Raman microscopy (also known as Raman microspectroscopy), is ideally suited to study variations of cellular composition on the scale of subcellular organelles, since its spatial resolution is as good as that of fluorescence microscopy. Both techniques exhibit the fingerprint sensitivity of vibrational spectroscopy toward biochemical composition, and can be used to follow a variety of cellular processes.

Molecular pathology via IR and Raman spectral imaging

“…Figure 2 Raman spectra of selected cellular components. Section 3.2) in Raman microscopy is similar to that of confocal fluorescence microscopy [22].…”

Evaluation of the impact of vitrification on the actin cytoskeleton of in vitro matured ovine oocytes by means of Raman microspectroscopy

“…It is possible to identify prominent spectral peaks in characteristic spectral regions. These features arise from the molecular vibration of components of the oocyte cortex such as lipids and proteins [27]. deformation of the proteins and lipids.…”

“…Raman offers the advantage of acquiring information regarding the biochemical constituents of the sample and their spatial distribution without labelling, on both fixed and unfixed cells. This technique has been, widely, used to study the biological changes of various cellular components during cell cycle progression, apoptosis, and response to stress [27]. On the other hand, studies focused on female gamete are restricted to a few works [28][29][30][31][32].…”

A micro-Raman spectroscopic investigation of leukemic U-937 cells treated with Crotalaria agatiflora Schweinf and the isolated compound madurensine

“…Two opposing interpretations have been used to discuss intensity changes of peaks during treatments. The first interpretation states that as the intensity of Raman peaks depends linearly on the concentration of molecules in a sample, a lower concentration will imply that the associated Raman peak decreases in intensity [13]. The second interpretation suggested that when molecules denature or lose their form and function the intensity of some Raman peaks will increase [7].…”