Rapid and Label-Free Histopathology of Oral Lesions Using Deep Learning Applied to Optical and Infrared Spectroscopic Imaging Data

Oral potentially malignant disorders (OPMDs) are precursors to over 80% of oral cancers. Hematoxylin and eosin (H&E) staining, followed by pathologist interpretation of tissue and cellular morphology, is the current gold standard for diagnosis. However, this method is qualitative, can result in errors during the multi-step diagnostic process, and results may have significant inter-observer variability. Chemical imaging (CI) offers a promising alternative, wherein label-free imaging is used to record both the morphology and the composition of tissue and artificial intelligence (AI) is used to objectively assign histologic information. Here, we employ quantum cascade laser (QCL)-based discrete frequency infrared (DFIR) chemical imaging to record data from oral tissues. In this proof-of-concept study, we focused on achieving tissue segmentation into three classes (connective tissue, dysplastic epithelium, and normal epithelium) using a convolutional neural network (CNN) applied to three bands of label-free DFIR data with paired darkfield visible imaging. Using pathologist-annotated H&E images as the ground truth, we demonstrate results that are 94.5% accurate with the ground truth using combined information from IR and darkfield microscopy in a deep learning framework. This chemical-imaging-based workflow for OPMD classification has the potential to enhance the efficiency and accuracy of clinical oral precancer diagnosis.

[1]  J. Cheville,et al.  Infrared spectroscopic laser scanning confocal microscopy for whole-slide chemical imaging , 2023, Nature communications.

[2]  O. Bouché,et al.  Deep learning for the prediction of the chemotherapy response of metastatic colorectal cancer: comparing and combining H&E staining histopathology and infrared spectral histopathology. , 2023, The Analyst.

[3]  R. Bhargava Digital Histopathology by Infrared Spectroscopic Imaging , 2023, Annual review of analytical chemistry.

[4]  J. Cheville,et al.  Deepfake Histologic Images for Enhancing Digital Pathology. , 2023, Laboratory investigation; a journal of technical methods and pathology.

[5]  A. Mosig,et al.  Fast and label-free automated detection of microsatellite status in early colon cancer using artificial intelligence integrated infrared imaging. , 2023, European journal of cancer.

[6]  R. Bhargava,et al.  Chemical imaging of cellular ultrastructure by null-deflection infrared spectroscopic measurements , 2022, Proceedings of the National Academy of Sciences of the United States of America.

[7]  S. Toyosawa,et al.  Oral squamous cell carcinoma diagnosis in digitized histological images using convolutional neural network , 2022, Journal of dental sciences.

[8]  Sumsum P. Sunny,et al.  Consensus guidelines on management of oral potentially malignant disorders , 2022, Indian journal of cancer.

[9]  J. Schwartz,et al.  A guide to nanoscale IR spectroscopy: resonance enhanced transduction in contact and tapping mode AFM-IR. , 2022, Chemical Society reviews.

[10]  R. Shaw,et al.  Prediction of malignant transformation in oral epithelial dysplasia using infrared absorbance spectra , 2022, PloS one.

[11]  S. Muller,et al.  Update from the 5th Edition of the World Health Organization Classification of Head and Neck Tumors: Tumours of the Oral Cavity and Mobile Tongue , 2022, Head and Neck Pathology.

[12]  Aaron C. Courville,et al.  Generative Adversarial Networks , 2022, 2023 14th International Conference on Computing Communication and Networking Technologies (ICCCNT).

[13]  E. Dartois,et al.  Photothermal AFM-IR spectroscopy and imaging: Status, challenges, and trends , 2022, Journal of Applied Physics.

[14]  Rong Wang,et al.  Oral Cancer Discrimination and Novel Oral Epithelial Dysplasia Stratification Using FTIR Imaging and Machine Learning , 2021, Diagnostics.

[15]  Ji‐Xin Cheng,et al.  Fluorescence-Detected Mid-Infrared Photothermal Microscopy. , 2021, Journal of the American Chemical Society.

[16]  D. Olinici,et al.  Molecular markers associated with potentially malignant oral lesions (Review) , 2021, Experimental and therapeutic medicine.

[17]  Ji‐Xin Cheng,et al.  Bond-selective imaging by optically sensing the mid-infrared photothermal effect , 2021, Science Advances.

[18]  Andreas C. Geiger,et al.  Fluorescence-Detected Mid-Infrared Photothermal Microscopy. , 2021, Journal of the American Chemical Society.

[19]  S. Hewitt,et al.  INFORM: INFrared-based ORganizational Measurements of tumor and its microenvironment to predict patient survival , 2021, Science Advances.

[20]  Jakob Nikolas Kather,et al.  Deep learning in cancer pathology: a new generation of clinical biomarkers , 2020, British Journal of Cancer.

[21]  A. King,et al.  Tumours of the oral cavity and pharynx , 2020 .

[22]  K. Ranganathan,et al.  Intra-observer and inter-observer variability in two grading systems for oral epithelial dysplasia: a multi-centre study in India. , 2020, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.

[23]  R. Zenobi,et al.  Infrared and Raman chemical imaging and spectroscopy at the nanoscale. , 2020, Chemical Society reviews.

[24]  C. Garnis,et al.  Oral potentially malignant disorders: A scoping review of prognostic biomarkers. , 2020, Critical reviews in oncology/hematology.

[25]  Dejun Chen,et al.  DFT-Calculated IR Spectrum Amide I, II, and III Band Contributions of N-Methylacetamide Fine Components , 2020, ACS omega.

[26]  R. Bhargava,et al.  All-digital histopathology by infrared-optical hybrid microscopy , 2020, Proceedings of the National Academy of Sciences.

[27]  E. D. Rekow,et al.  Digital dentistry: The new state of the art - Is it disruptive or destructive? , 2020, Dental materials : official publication of the Academy of Dental Materials.

[28]  Rohit Bhargava,et al.  Multicolor Discrete Frequency Infrared Spectroscopic Imaging. , 2019, Analytical chemistry.

[29]  Andre Kajdacsy-Balla,et al.  Simultaneous cancer and tumor microenvironment subtyping using confocal infrared microscopy for all-digital molecular histopathology , 2018, Proceedings of the National Academy of Sciences.

[30]  Nicholas Stone,et al.  Mid-IR hyperspectral imaging for label-free histopathology and cytology , 2018 .

[31]  U. Hegde,et al.  Inter- and Intra-Observer Variability in Diagnosis of Oral Dysplasia , 2017, Asian Pacific journal of cancer prevention : APJCP.

[32]  George Papandreou,et al.  Rethinking Atrous Convolution for Semantic Image Segmentation , 2017, ArXiv.

[33]  Ihtesham ur Rehman,et al.  Advances in Fourier transform infrared (FTIR) spectroscopy of biological tissues , 2017 .

[34]  F. Nikolajeff,et al.  Role of Infrared Spectroscopy and Imaging in Cancer Diagnosis. , 2017, Current medicinal chemistry.

[35]  Alexandre Dazzi,et al.  AFM-IR: Technology and Applications in Nanoscale Infrared Spectroscopy and Chemical Imaging. , 2017, Chemical reviews.

[36]  Delong Zhang,et al.  Depth-resolved mid-infrared photothermal imaging of living cells and organisms with submicrometer spatial resolution , 2016, Science Advances.

[37]  Rohit Bhargava,et al.  Development of a practical spatial-spectral analysis protocol for breast histopathology using Fourier transform infrared spectroscopic imaging. , 2016, Faraday discussions.

[38]  Peter Gardner,et al.  Fundamental developments in infrared spectroscopic imaging for biomedical applications. , 2016, Chemical Society reviews.

[39]  A. Ariyawardana,et al.  Malignant transformation of oral leukoplakia: a systematic review of observational studies. , 2016, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.

[40]  Jian Sun,et al.  Deep Residual Learning for Image Recognition , 2015, 2016 IEEE Conference on Computer Vision and Pattern Recognition (CVPR).

[41]  J. McDevitt,et al.  Interobserver agreement in dysplasia grading: toward an enhanced gold standard for clinical pathology trials. , 2015, Oral surgery, oral medicine, oral pathology and oral radiology.

[42]  David Mayerich,et al.  Stain-less staining for computed histopathology. , 2015, Technology.

[43]  G. Lloyd,et al.  Infrared micro-spectroscopy for cyto-pathological classification of esophageal cells. , 2015, The Analyst.

[44]  Trevor Darrell,et al.  Fully convolutional networks for semantic segmentation , 2014, 2015 IEEE Conference on Computer Vision and Pattern Recognition (CVPR).

[45]  Rohit Bhargava,et al.  Using Fourier transform IR spectroscopy to analyze biological materials , 2014, Nature Protocols.

[46]  Antonella I. Mazur,et al.  Molecular pathology via IR and Raman spectral imaging , 2013, Journal of biophotonics.

[47]  L. Miller,et al.  From structure to cellular mechanism with infrared microspectroscopy. , 2010, Current opinion in structural biology.

[48]  S. Kamel‐Reid,et al.  Abnormal DNA content in oral epithelial dysplasia is associated with increased risk of progression to carcinoma , 2010, British Journal of Cancer.

[49]  A. Jara-Lazaro,et al.  Digital pathology: exploring its applications in diagnostic surgical pathology practice , 2010, Pathology.

[50]  T. Aach,et al.  Toward a multimodal cell analysis of brush biopsies for the early detection of oral cancer , 2009, Cancer.

[51]  S. Mukamel,et al.  Two-Dimensional Vibrational Lineshapes of Amide III, II, I and A Bands in a Helical Peptide. , 2008, Journal of molecular liquids.

[52]  S. Warnakulasuriya,et al.  Nomenclature and classification of potentially malignant disorders of the oral mucosa. , 2007, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.

[53]  Philip Sloan,et al.  Why oral histopathology suffers inter-observer variability on grading oral epithelial dysplasia: an attempt to understand the sources of variation. , 2007, Oral oncology.

[54]  Philip Sloan,et al.  Evaluation of a new binary system of grading oral epithelial dysplasia for prediction of malignant transformation. , 2006, Oral oncology.

[55]  Cyril Petibois,et al.  Chemical mapping of tumor progression by FT-IR imaging: towards molecular histopathology. , 2006, Trends in biotechnology.

[56]  S. Kazarian,et al.  Applications of ATR-FTIR spectroscopic imaging to biomedical samples. , 2006, Biochimica et biophysica acta.

[57]  I. W. Levin,et al.  Fourier transform infrared vibrational spectroscopic imaging: integrating microscopy and molecular recognition. , 2005, Annual review of physical chemistry.

[58]  M. Gaigeot,et al.  Infrared Spectroscopy of N-Methylacetamide Revisited by ab Initio Molecular Dynamics Simulations. , 2005, Journal of chemical theory and computation.

[59]  S. Hewitt,et al.  Infrared spectroscopic imaging for histopathologic recognition , 2005, Nature Biotechnology.

[60]  S. Cai,et al.  Identification of beta-turn and random coil amide III infrared bands for secondary structure estimation of proteins. , 1999, Biophysical chemistry.

[61]  S. Tanaka,et al.  Quantitative Estimation of α-Helix Coil Content in Bovine Serum Albumin by Fourier Transform-Infrared Spectroscopy , 1987 .

[62]  R. Mendelsohn,et al.  Thermal denaturation of globular proteins. Fourier transform-infrared studies of the amide III spectral region. , 1987, Biophysical journal.

[63]  Mitsuo Tasumi,et al.  Infrared studies of the less stable cis form of N-methylformmaide and N-methylacetamide in low-temperature nitrogen matrices and vibrational analyses of the trans and cis forms of these molecules , 1984 .

[64]  Richard L Zimmermann,et al.  The Impact of Technological Innovation on Dentistry. , 2023, Advances in experimental medicine and biology.

[65]  Guigang Zhang,et al.  Deep Learning , 2016, Int. J. Semantic Comput..