Issue 14, 2013

Substrate contributions in micro-ATR of thin samples: implications for analysis of cells, tissue and biological fluids

Abstract

Low-e microscope slides are a common substrate for biological samples. Typically they are used for transflection infrared microspectroscopy but increasingly they are also being used for micro-ATR experiments since it is assumed that the FTIR-ATR absorbance spectra of cells and tissue on low-e substrates will not contain any spectral contributions from the substrate materials. This, in part, is due to the expectation that all the infrared light will be reflected at the highly reflective surface. At low sample thicknesses, however (e.g. less than 2 μm) the electric field does indeed penetrate through the substrate layers and undergoes absorption, from the glass supporting layer making up the majority of the slide. In this paper we show experimental evidence of the substrate contributions in ATR spectra and also a theoretical model giving insight into the spectral contributions of the substrate as a function of sample thickness.

Graphical abstract: Substrate contributions in micro-ATR of thin samples: implications for analysis of cells, tissue and biological fluids

Article information

Article type
Paper
Submitted
20 Feb 2013
Accepted
31 May 2013
First published
31 May 2013

Analyst, 2013,138, 4139-4146

Substrate contributions in micro-ATR of thin samples: implications for analysis of cells, tissue and biological fluids

P. Bassan, A. Sachdeva, J. Lee and P. Gardner, Analyst, 2013, 138, 4139 DOI: 10.1039/C3AN00363A

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