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Optimisation of sample preparation using glass slides for spectral pathology

Dowling, LM; Roach, P; Rutter, A; Yousef, I; Pillai, S; Latham, D; van Pittius, DG; Sule-Suso, J


LM Dowling

P Roach

I Yousef

S Pillai

D Latham

DG van Pittius


The clinical translation of Fourier Transform Infrared (FTIR) 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 FTIR microspectroscopy and pathology. This should entail minimal disruption of standard clinical practice while achieving good quality FTIR spectral data. In fact, the recently described possibility of obtaining FTIR spectra of cells placed on glass substrates brings FTIR 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 FTIR 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.

Journal Article Type Article
Acceptance Date Jul 1, 2020
Publication Date Jul 14, 2020
Journal Applied Spectroscopy
Print ISSN 0003-7028
Publisher SAGE Publications
Peer Reviewed Peer Reviewed
Volume 75
Issue 3
Pages 343-350
Keywords FTIR microspectroscopy, lung cancer, glass coverslips, cell fixation, cytospin, smear
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