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. 2021 Apr 28;12(5):3008-3020.
doi: 10.1364/BOE.420882. eCollection 2021 May 1.

Raman spectroscopy for guidance of vulvar cancer surgery: a pilot study

Helena C van Doorn  1   2 , Elisa M Barroso  3   2 , Senada Koljenović  3 , Patricia C Ewing-Graham  3 , M Rosa N Soares  3 , Nick J van de Berg  1 , Tom C Bakker Schut  4 , Gerwin J Puppels  4
Affiliations

Raman spectroscopy for guidance of vulvar cancer surgery: a pilot study

Helena C van Doorn et al. Biomed Opt Express. .

Abstract

For vulvar squamous cell carcinoma (VSCC), the mainstay of treatment is surgical removal with tumour-free margins. Surgeons still operate without objective tools that provide margin-status. This study assesses Raman spectroscopy potentiality for distinguishing ex-vivo VSCC from healthy tissue in 11 patients. Grid-based Raman maps were obtained from processed spectra. Water content and C-H band ratio (2,910-2,966 cm-1 / 2810-2890 cm-1) were calculated per spectrum and used as linear discriminant parameters. Healthy tissue was differentiated from VSCC with 0.90 discriminative power, 0.79 sensitivity and 0.86 specificity.This is an important step towards the development of objective tools for VSCC surgical guidance.

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Conflict of interest statement

S. Koljenović, and G.J. Puppels have ownership interest (including patents) in RiverD International BV. T.C. Bakker Schut, M. Rosa N. Soares are employees of RiverD International BV. M. Rosa N. Soares is also employee of RiverD International BV. G.J. Puppels is employed with RiverD International BV as a CTO & Managing Director. No potential conflicts of interest were disclosed by the other authors.

Figures

Fig. 1.
Fig. 1.
Overview of the experimental protocol. A. Immediately after surgical resection, the specimen was brought by the surgeon to the pathology room, and the anatomical orientation in relation to midline structures was digitally recorded (‘L’ means left). B. Raman tissue section (‘Raman exp.’) was cut perpendicular to the resection surface. C. The Raman tissue section was inserted into a cartridge. The area to be measured, as defined by the surgeon and/or pathologist, containing tumour and surrounding healthy tissue. D. After performing Raman mapping experiments, the specimen was routinely processed, and a hematoxylin and eosin slide was made from the whole Raman tissue section measured. Histopathological annotation of tumour (T), healthy tissue (H), and of the tumour border (red line) was performed.
Fig. 2.
Fig. 2.
Examples of HCA results for 3 Raman tissue sections from 3 patients (Column A-C). 1. Macroscopic photograph after insertion in the cartridge. 2. Cluster map color-coded according to the color of each cluster average (Column D). 3. Hematoxylin and eosin (H&E) stained section obtained from the measured Raman tissue section surface, with tumour border (red line), tumour (T), and healthy surrounding tissue (H) indicated by the pathologist. D. The 20 cluster averages (c1-c20) resulted from the hierarchical clustering analysis: 14 cluster averages were histopatholgically correlated to a specific tissue structure (‘Cluster averages’ graph) and 6 cluster averages were considered outliers (‘Cluster averages: outliers’ graph).
Fig. 3.
Fig. 3.
Histograms of absolute frequencies for C-H band ratio (A and B) and water content (C and D) of healthy tissue spectra (adipose tissue, fibrocollagenous tissue, blood vessels, and fibromuscular stroma) and of tumour spectra.
Fig. 4.
Fig. 4.
ROC curve for leave one patient out validation of the LDA VSCC discrimination model based on water percentage and C-H band ratio.
Fig. 5.
Fig. 5.
Two examples (A and B) of Raman mapping experiments that were excluded. A. Macroscopic photograph after insertion in the cartridge with corresponding hematoxylin and eosin (H&E) slide. The histopathological annotation of tumour (T), healthy tissue (H), and of the tumour border (red line) is performed in the H&E slide. There is not a complete overlap between the shape of the Raman tissue section (Macroscopy) and the respective H&E-stained section (Histology). B. Macroscopic photograph after insertion in the cartridge with corresponding hematoxylin and eosin (H&E) slide. The histopathological annotation of healthy tissue (H) is performed in the H&E slide. The Raman tissue section does not contain tumour.
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References

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