Raman Spectroscopy of the Hematoxylin - Eosin Stained Tissue

Luisa Andronia, Vioara Mireșan, Aurelia Coroian, Ioana Pop, Camelia Răducu, Ancuța Rotaru, Daniel Cocan, Simona Cîntă Pânzaru, Ioan Domșa, Cristian Ovidiu Coroian

Abstract


Micro-Raman spectra of the hematoxylin-eosin stained tissue have been recorded and analyzed for both normal and carcinoma colon samples from nine patients in order to evaluate the compatibility of H&E staining with Raman spectroscopy of tissue. An additional aim was to evaluate if the “normal†tissue from various geographical locations is the same in terms of Raman signal from equivalent samples reported in the literature.  FT-Raman signal of the formalin -fixed tissue was used for comparison. The overall Raman signal of stained tissue is a superposition of Raman scattering intensity from staining agents, tissue and auto-fluorescence of the tissue and staining agents. Due to the different Raman cross section of the species, the superposition giving the overall Raman spectral shape is not a linear combination, i.e. subtracting the Raman signal of staining agents does not necessarily result in the Raman signal of unstained tissue. Moreover, the H&E staining agents showed less influence on the overall tissue signal, allowing identify specific Raman bands of tissue. The major influence on the overall signal was produced by the resins (biomount, xylene), which are not compatible with the tissue Raman diagnostic. Vibrational analysis results of the H&E stained tissue samples point out the capability of the Raman techniques to evaluate the enormous archival tissue material available in clinical diagnostic laboratories and could further better assist the histopathology conclusions with high specificity and sensitivity.


Keywords


Micro-Raman, colon carcinoma, adenocarcinoma, hematoxylin, eosin, stained tissue.

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