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Optimization of sample preparation using glass slides for spectral pathology

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journal contribution
posted on 23.03.2021, 11:46 by Lewis M. Dowling, Paul RoachPaul Roach, Abigail V. Rutter, Ibraheem Yousef, Srinivas Pilla, Deborah Latham, Daniel G. van Pittius, Josep Sulé-Suso
The clinical translation of Fourier transform infrared (FT-IR) microspectroscopy in pathology will require bringing this technique as close as possible to standard practice in pathology departments. An important step is sample preparation for both FT-IR microspectroscopy and pathology. This should entail minimal disruption of standard clinical practice while achieving good quality FT-IR spectral data. In fact, the recently described possibility of obtaining FT-IR spectra of cells placed on glass substrates brings FT-IR microspectroscopy closer to a clinical application. We have now furthered this work in order to identify two different types of lung cancer cells placed on glass coverslips. Two types of sample preparation which are widely used in pathology, cytospin and smear, have been used. Samples were fixed with either methanol, used in pathology, or formalin (4% paraformaldehyde) used widely in spectroscopy. Fixation with methanol (alcohol-based fixative) removed lipids from cells causing a decrease in intensity of the peaks at 2850 cm−1 and 2920 cm−1. Nevertheless, we show for the first time that using either type of sample preparation and fixation on thin glass coverslips allowed to differentiate between two different types of lung cancer cells using either the lipid region or the fingerprint region ranging from 1800 cm−1 to 1350 cm−1. We believe that formalin-fixed cytospin samples would be preferred to study cells on thin coverslips using FT-IR microspectroscopy. This work presents a clear indication for future advances in clinical assessment of samples within pathology units to gain a deeper understanding of cells/tissues under investigation.

Funding

Cancer Centre and University Hospitals of North Midlands charities

Alba Synchrotron Light Source

History

School

  • Science

Department

  • Chemistry

Published in

Applied Spectroscopy

Volume

75

Issue

3

Pages

343 - 350

Publisher

SAGE Publications

Version

VoR (Version of Record)

Rights holder

© The Authors

Publisher statement

This is an Open Access Article. It is published by SAGE under the Creative Commons Attribution-NonCommercial 4.0 International Licence (CC BY-NC 4.0). Full details of this licence are available at: https://creativecommons.org/licenses/by-nc/4.0/

Acceptance date

30/06/2020

Publication date

2020-07-14

Copyright date

2020

ISSN

0003-7028

eISSN

1943-3530

Language

en

Depositor

Dr Paul Roach. Deposit date: 18 March 2021