Mode conversion of qOBM (quantitative oblique back-illumination microscopy) stain-free tissue images to emulate H&E histology via deep learning

Tanishq Mathew Abraham; Paloma Casteleiro Costa; Caroline Filan; Francisco E. Robles; Richard Levenson

doi:10.1117/12.2649484

16 March 2023 Mode conversion of qOBM (quantitative oblique back-illumination microscopy) stain-free tissue images to emulate H&E histology via deep learning

Tanishq Mathew Abraham, Paloma Casteleiro Costa, Caroline Filan, Francisco E. Robles, Richard Levenson

Author Affiliations +

Proceedings Volume 12391, Label-free Biomedical Imaging and Sensing (LBIS) 2023; 1239109 (2023) https://doi.org/10.1117/12.2649484
Event: SPIE BiOS, 2023, San Francisco, California, United States

Abstract

Slide-free microscopy techniques have been proposed for accelerating standard histopathology and intraoperative guidance. One such technology is quantitative oblique back-illumination microscopy (qOBM), which enables real-time, label-free quantitative phase imaging of thick, unsectioned in-vivo and ex-vivo tissues. However, the grayscale phase contrast provided by qOBM differs from the colored histology images familiar to pathologists and clinicians, limiting its current potential for adoption. Here we demonstrate the application of unsupervised deep learning using a Cycleconsistent Generative Adversarial Network (CycleGAN) model to transform qOBM images into virtual hematoxylin and eosin (H&E)-stained images. The models were trained on a dataset of qOBM and H&E images of similar regions in excised brain tissue from a 9 L gliosarcoma rat tumor model. We observed successful qOBM-to-H&E conversion of both uninvolved and tumor-containing specimens, as demonstrated by a classifier test. We describe several crucial preprocessing steps that improve the quality of conversion, such as intensity inversion, pixel harmonization, and color normalization. This unsupervised deep learning framework does exhibit occasional subpar performance; for example, as with GANs in general, it can create so-called “hallucinations”, displaying features not actually present in the original qOBM images. We anticipate that this behavior can be minimized with more extensive training and deployment of advanced ML techniques, and that virtual-H&E-converted qOBM imaging will prove safe and appropriate for rapid tissue imaging applications.

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Tanishq Mathew Abraham, Paloma Casteleiro Costa, Caroline Filan, Francisco E. Robles, and Richard Levenson "Mode conversion of qOBM (quantitative oblique back-illumination microscopy) stain-free tissue images to emulate H&E histology via deep learning", Proc. SPIE 12391, Label-free Biomedical Imaging and Sensing (LBIS) 2023, 1239109 (16 March 2023); https://doi.org/10.1117/12.2649484

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