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Comparative Study
. 2006 May 22;94(10):1460-4.
doi: 10.1038/sj.bjc.6603102.

Raman spectroscopy: elucidation of biochemical changes in carcinogenesis of oesophagus

G Shetty  1 , C Kendall, N Shepherd, N Stone, H Barr
Affiliations
Comparative Study

Raman spectroscopy: elucidation of biochemical changes in carcinogenesis of oesophagus

G Shetty et al. Br J Cancer. .

Abstract

Several techniques are under development to diagnose oesophageal adenocarcinoma at an earlier stage. We have demonstrated the potential of Raman spectroscopy, an optical diagnostic technique, for the identification and classification of malignant changes. However, there is no clear recognition of the biochemical changes that distinguish between the different stages of disease. Our aim is to understand these changes through Raman mapping studies. Raman spectral mapping was used to analyse 20-microm sections of tissue from 29 snap-frozen oesophageal biopsies. Contiguous haematoxylin and eosin sections were reviewed by a consultant pathologist. Principal component analysis was used to identify the major differences between the spectra across each map. Pseudocolour score maps were generated and the peaks of corresponding loads identified enabling visualisation of the biochemical changes associated with malignancy. Changes were noted in the distribution of DNA, glycogen, lipids and proteins. The mean spectra obtained from selected regions demonstrate increased levels of glycogen in the squamous area compared with increased DNA levels in the abnormal region. Raman spectroscopy is a highly sensitive and specific technique for demonstration of biochemical changes in the carcinogenesis of Barrett's oesophagus. There is potential for in vivo application for real-time endoscopic optical diagnosis.

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Figures

Figure 1
Figure 1
Haematoxylin and eosin stained sections with pathology labelled. (A) Regions 1 and 2 normal squamous and regions 3 and 4 HGD. (B) Region 1 normal squamous and regions 2, 3 and 4 adenocarcinoma.
Figure 2
Figure 2
First eight pseudocolour principal component score maps and corresponding loads for the normal squamous and HGD sample (H&E stained section Figure 1A). PC 1 top left to PC 8 bottom right.
Figure 3
Figure 3
Normal squamous and HGD sample (H&E stained section Figure 1A). (A) Third principal component score map with selected regions marked. (B) Mean Raman spectra from selected regions. (C) Relative concentration of selected biochemical constituents calculated with mean spectra from selected regions. Normal squamous and adenocarcinoma sample (H&E stained section Figure 1B). (D) Third principal component score map with selected regions marked. (E) Mean Raman spectra from selected regions. (F) Relative concentration of selected biochemical constituents calculated with mean spectra from selected regions.
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