Reflection contributions to the dispersion artefact in FTIR spectra of single biological cells

Paul Bassan; Hugh J. Byrne; Joe Lee; Franck Bonnier; Colin Clarke; Paul Dumas; Ehsan Gazi; Michael D. Brown; Noel W. Clarke; Peter Gardner

doi:10.1039/B821349F

Reflection contributions to the dispersion artefact in FTIR spectra of single biological cells†

Paul Bassan,^a Hugh J. Byrne,*^b Joe Lee,^a Franck Bonnier,^b Colin Clarke,^b Paul Dumas,^c Ehsan Gazi,^d Michael D. Brown,^d Noel W. Clarke^def and Peter Gardner^a

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* Corresponding authors

^a School of Chemical Engineering and Analytical Science, Manchester Interdisciplinary Biocentre, The University of Manchester, 131 Princess Street, Manchester, UK
E-mail: Peter.Gardner@Manchester.ac.uk

^b Focas Research Institute, Dublin Institute of Technology, Kevin Street, Dublin 8, Ireland

^c Synchrotron SOLEIL, L'Orme des Merisiers, BP48 - Saint Aubin, 91192 Gif-sur-Yvette Cedex, France

^d Genito Urinary Cancer Research Group, School of Cancer and Imaging Sciences Paterson Institute for Cancer Research, University of Manchester, Wilmslow Road, Manchester, UK

^e Department of Urology, The Christie NHS Foundation Trust, Manchester, UK

^f Department of Urology, Salford Royal NHS Foundation Trust, Salford, UK

Abstract

Fourier transform infrared spectra of a single cell in transflection geometry are seen to vary significantly with position on the cell, showing a distorted derivative-like lineshape in the region of the optically dense nucleus. A similar behaviour is observable in a model system of the protein albumin doped in a potassium bromide disk. It is demonstrated that the spectrum at any point is a weighted sum of the sample reflection and transmission and that the dominance of the reflection spectrum in optically dense regions can account for some of the spectral distortions previously attributed to dispersion artefacts. Rather than being an artefact, the reflection contribution is ever present in transflection spectra and it is further demonstrated that the reflection characteristics can be used for cellular mapping.

This article is part of the themed collection: Optical Diagnosis

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Reflection contributions to the dispersion artefact in FTIR spectra of single biological cells†

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Reflection contributions to the dispersion artefact in FTIR spectra of single biological cells

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