Theranostics 2019; 9(9):2541-2554. doi:10.7150/thno.32655

Research Paper

Rapid histology of laryngeal squamous cell carcinoma with deep-learning based stimulated Raman scattering microscopy

Lili Zhang1,2#, Yongzheng Wu1#, Bin Zheng3#✉, Lizhong Su3, Yuan Chen4, Shuang Ma4, Qinqin Hu4, Xiang Zou5, Lie Yao5, Yinlong Yang6, Liang Chen5, Ying Mao5, Yan Chen1✉, Minbiao Ji1,2✉

1. State Key Laboratory of Surface Physics and Department of Physics, Fudan University, Shanghai 200433, China
2. Human Phenome Institute, Multiscale Research Institute of Complex Systems, Key Laboratory of Micro and Nano Photonic Structures (Ministry of Education), Fudan University, Shanghai 200433, China
3. Department of Otolaryngology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China
4. Department of Pathology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China
5. Department of Neurosurgery, Department of Pancreatic Surgery, Huashan Hospital, Fudan University, Shanghai 200040, China
6. Department of Breast Surgery, Fudan University Shanghai Cancer Center; Department of Oncology, Shanghai Medical College; Fudan University, Shanghai 200040, China
# These authors contributed equally.

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Citation:
Zhang L, Wu Y, Zheng B, Su L, Chen Y, Ma S, Hu Q, Zou X, Yao L, Yang Y, Chen L, Mao Y, Chen Y, Ji M. Rapid histology of laryngeal squamous cell carcinoma with deep-learning based stimulated Raman scattering microscopy. Theranostics 2019; 9(9):2541-2554. doi:10.7150/thno.32655. Available from http://www.thno.org/v09p2541.htm

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Abstract

Maximal resection of tumor while preserving the adjacent healthy tissue is particularly important for larynx surgery, hence precise and rapid intraoperative histology of laryngeal tissue is crucial for providing optimal surgical outcomes. We hypothesized that deep-learning based stimulated Raman scattering (SRS) microscopy could provide automated and accurate diagnosis of laryngeal squamous cell carcinoma on fresh, unprocessed surgical specimens without fixation, sectioning or staining.

Methods: We first compared 80 pairs of adjacent frozen sections imaged with SRS and standard hematoxylin and eosin histology to evaluate their concordance. We then applied SRS imaging on fresh surgical tissues from 45 patients to reveal key diagnostic features, based on which we have constructed a deep learning based model to generate automated histologic results. 18,750 SRS fields of views were used to train and cross-validate our 34-layered residual convolutional neural network, which was used to classify 33 untrained fresh larynx surgical samples into normal and neoplasia. Furthermore, we simulated intraoperative evaluation of resection margins on totally removed larynxes.

Results: We demonstrated near-perfect diagnostic concordance (Cohen's kappa, κ > 0.90) between SRS and standard histology as evaluated by three pathologists. And deep-learning based SRS correctly classified 33 independent surgical specimens with 100% accuracy. We also demonstrated that our method could identify tissue neoplasia at the simulated resection margins that appear grossly normal with naked eyes.

Conclusion: Our results indicated that SRS histology integrated with deep learning algorithm provides potential for delivering rapid intraoperative diagnosis that could aid the surgical management of laryngeal cancer.

Keywords: label-free imaging, stimulated Raman scattering, intraoperative histology, laryngeal cancer, head and neck