Stain-free Histopathology of Basal Cell Carcinoma by Dual Vibration Resonance Frequency CARS Microscopy

Pathology Oncology Research : POR
Norbert KissNorbert Wikonkál

Abstract

Basal cell carcinoma (BCC) is the most common malignancy in Caucasians. Nonlinear microscopy has been previously utilized for the imaging of BCC, but the captured images do not correlate with H&E staining. Recently, Freudiger et al. introduced a novel method to visualize tissue morphology analogous to H&E staining, using coherent anti-Stokes Raman scattering (CARS) technique. In our present work, we introduce a novel algorithm to post-process images obtained from dual vibration resonance frequency (DVRF) CARS measurements to acquire high-quality pseudo H&E images of BCC samples. We adapted our CARS setup to utilize the distinct vibrational properties of CH3 (mainly in proteins) and CH2 bonds (primarily in lipids). In a narrowband setup, the central wavelength of the pump laser is set to 791 nm and 796 nm to obtain optimal excitation. Due to the partial overlap of the excitation spectra and the 5-10 nm FWHM spectral bandwidth of our lasers, we set the wavelengths to 790 nm (proteins) and 800 nm (lipids). Nonresonant background from water molecules also reduces the chemical selectivity which can be significantly improved if we subtract the DVRF images from each other. As a result, we acquired two images: one for "lipids" and one ...Continue Reading

References

Jul 22, 2010·Journal of Biophotonics·Nadine VoglerJürgen Popp
Jul 11, 2012·Skin Research and Technology : Official Journal of International Society for Bioengineering and the Skin (ISBS) [and] International Society for Digital Imaging of Skin (ISDIS) [and] International Society for Skin Imaging (ISSI)·Stefania SeidenariKarsten König
Aug 22, 2012·Laboratory Investigation; a Journal of Technical Methods and Pathology·Christian W FreudigerGeoffrey S Young
Apr 16, 2014·American Journal of Clinical Dermatology·Charlotte M ClarkJulian M Mackay-Wiggan
Nov 25, 2016·Scientific Reports·Alex Soares DuartePhilipp Kukura
Nov 30, 2016·Biomedical Optics Express·Dóra HaluszkaNorbert M Wikonkál

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Citations

Jan 15, 2021·Chemistry, an Asian Journal·Adukkadan N RamyaKaustabh K Maiti
Aug 28, 2021·Diagnostics·Szabolcs BozsányiNorbert M Wikonkál

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Methods Mentioned

BETA
biopsy

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ImageJ

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