Fast stimulated Raman and second harmonic generation imaging for intraoperative gastro-intestinal cancer detection

Barbara Sarri,Didier Marguet,Emma Simon,Cécile Cador,Flora Poizat,Julien Wojak,Fabrice Caillol,Xavier Audier,Marc Giovannini,Rafaël Canonge,Hervé Rigneault

doi:10.1038/s41598-019-46489-x

Abstract

Conventional haematoxylin, eosin and saffron (HES) histopathology, currently the ‘gold-standard’ for pathological diagnosis of cancer, requires extensive sample preparations that are achieved within time scales that are not compatible with intra-operative situations where quick decisions must be taken. Providing to pathologists a close to real-time technology revealing tissue structures at the cellular level with HES histologic quality would provide an invaluable tool for surgery guidance with evident clinical benefit. Here, we specifically develop a stimulated Raman imaging based framework that demonstrates gastro-intestinal (GI) cancer detection of unprocessed human surgical specimens. The generated stimulated Raman histology (SRH) images combine chemical and collagen information to mimic conventional HES histopathology staining. We report excellent agreements between SRH and HES images acquire on the same patients for healthy, pre-cancerous and cancerous colon and pancreas tissue sections. We also develop a novel fast SRH imaging modality that captures at the pixel level all the information necessary to provide instantaneous SRH images. These developments pave the way for instantaneous label free GI histology in an intra-operative context.

Highlights

Most organs from the gastrointestinal tract are subjected to cancer development
In order to speed up the stimulated Raman histology (SRH) acquisition time we have demonstrated a novel imaging platform, FM-stimulated Raman scattering (SRS), able to perform SRH images of GI tissues over millimeters large areas sections in a time that is compatible with the operatory room workflow
In this paper, stimulated Raman scattering (SRS) microscopy was used in association with second harmonic generation (SHG) microscopy to visualize tissue sections from healthy and cancerous human gastro-intestinal tract

Summary

Introduction

Most organs from the gastrointestinal tract are subjected to cancer development. Nowadays, colorectal cancer is the fourth cause of cancer related-death worldwide and pancreas cancers are fatal with a survival rate of only 1–5% after 5 years[1,2,3]. Most importantly the unique ability of SRS to image CH2 and CH3 chemical bonds found predominantly in cell bodies and cell nuclei, respectively, was shown to provide rapid label free microscopic images of healthy and cancer brain sections in near-perfect concordance with standard histology[12,14]. This major step forward opens the completely new field of stimulated Raman histology (SRH) for real-time label-free intra-operative surgical cancer tissue resections and treatments[15]. It paves the way to use SRH for GI tissues cancer identification as a possible alternative to extemporaneous histology to provide accurate HES like images to assist in surgical decision making, in an intra-operative context

Methods

Results

Discussion

Conclusion