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The inherent problem of transflection-mode infrared spectroscopic microscopy and the ramifications for biomedical single point and imaging applications.

Journal article
Authors Paul Bassan
Joe Lee
Ashwin Sachdeva
Juliana Pissardini
Konrad M Dorling
John S. Fletcher
Alex Henderson
Peter Gardner
Published in The Analyst
Volume 138
Issue 1
Pages 144-57
ISSN 1364-5528
Publication year 2013
Published at Department of Chemistry and Molecular Biology
Pages 144-57
Language en
Links dx.doi.org/10.1039/c2an36090j
Subject categories Analytical Chemistry

Abstract

Transflection-mode FTIR spectroscopy has become a popular method of measuring spectra from biomedical and other samples due to the relative low cost of substrates compared to transmission windows, and a higher absorbance due to a double pass through the same sample approximately doubling the effective path length. In this publication we state an optical description of samples on multilayer low-e reflective substrates. Using this model we are able to explain in detail the so-called electric-field standing wave effect and rationalise the non-linear change in absorbance with sample thickness. The ramifications of this non-linear change, for imaging and classification systems, where a model is built from tissue sectioned at a particular thickness and compared with tissue of a different thickness are discussed. We show that spectra can be distorted such that classification fails leading to inaccurate tissue segmentation which may have subsequent implications for disease diagnostics applications.

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