Effect of substrate choice and tissue type on tissue preparation for spectral histopathology by Raman microspectroscopy

Leanne M. Fullwood, Dave Griffiths, Katherine Ashton, Timothy P. Dawson, Robert W. Lea, Charles Davis, Franck Bonnier, Hugh J. Byrne, Matthew J. Baker

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Raman spectroscopy is a non-destructive, non-invasive, rapid and economical technique which has the potential to be an excellent method for the diagnosis of cancer and understanding disease progression through retrospective studies of archived tissue samples. Historically, biobanks are generally comprised of formalin fixed paraffin preserved tissue and as a result these specimens are often used in spectroscopic research. Tissue in this state has to be dewaxed prior to Raman analysis to reduce paraffin contributions in the spectra. However, although the procedures are derived from histopathological clinical practice, the efficacy of the dewaxing procedures that are currently employed is questionable. Ineffective removal of paraffin results in corruption of the spectra and previous experiments have shown that the efficacy can depend on the dewaxing medium and processing time. The aim of this study was to investigate the influence of commonly used spectroscopic substrates (CaF2, Spectrosil quartz and low-E slides) and the influence of different histological tissue types (normal, cancerous and metastatic) on tissue preparation and to assess their use for spectral histopathology. Results show that CaF2 followed by Spectrosil contribute the least to the spectral background. However, both substrates retain paraffin after dewaxing. Low-E substrates, which exhibit the most intense spectral background, do not retain wax and resulting spectra are not affected by paraffin peaks. We also show a disparity in paraffin retention depending upon the histological identity of the tissue with abnormal tissue retaining more paraffin than normal.

LanguageEnglish
Pages446-454
Number of pages9
JournalAnalyst
Volume139
Issue number2
Early online date25 Nov 2013
DOIs
Publication statusPublished - 21 Jan 2014

Fingerprint

histopathology
Paraffin
Paraffins
Tissue
substrate
Dewaxing
Substrates
Paraffin waxes
Quartz
Raman Spectrum Analysis
Waxes
corruption
Raman spectroscopy
wax
effect
tissue
Formaldehyde
Disease Progression
cancer
Retrospective Studies

Keywords

  • tissue preparation
  • spectral histopathology
  • Raman microspectroscopy

Cite this

Fullwood, Leanne M. ; Griffiths, Dave ; Ashton, Katherine ; Dawson, Timothy P. ; Lea, Robert W. ; Davis, Charles ; Bonnier, Franck ; Byrne, Hugh J. ; Baker, Matthew J. / Effect of substrate choice and tissue type on tissue preparation for spectral histopathology by Raman microspectroscopy. In: Analyst. 2014 ; Vol. 139, No. 2. pp. 446-454.
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Fullwood, LM, Griffiths, D, Ashton, K, Dawson, TP, Lea, RW, Davis, C, Bonnier, F, Byrne, HJ & Baker, MJ 2014, 'Effect of substrate choice and tissue type on tissue preparation for spectral histopathology by Raman microspectroscopy' Analyst, vol. 139, no. 2, pp. 446-454. https://doi.org/10.1039/c3an01832f

Effect of substrate choice and tissue type on tissue preparation for spectral histopathology by Raman microspectroscopy. / Fullwood, Leanne M.; Griffiths, Dave; Ashton, Katherine; Dawson, Timothy P.; Lea, Robert W.; Davis, Charles; Bonnier, Franck; Byrne, Hugh J.; Baker, Matthew J.

In: Analyst, Vol. 139, No. 2, 21.01.2014, p. 446-454.

Research output: Contribution to journalArticle

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