Shogo Shinohara1, Kazuo Funabiki2, Masahiro Kikuchi3, Shinji Takebayashi4, Kiyomi Hamaguchi4, Shigeo Hara5, Daisuke Yamashita5, Yukihiro Imai6, Akira Mizoguchi7. 1. Department of Otolaryngology-Head and Neck Surgery, Kobe City Medical Center General Hospital, Minatojima-Minamimachi 2-1-1, Chuo-ku, Kobe 650-0047, Japan. Electronic address: sinosino@kcho.jp. 2. Foundation for Biomedical Research and Innovation at Kobe, 1-5-4 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan. 3. Department of Otolaryngology-Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Sakyo-Ku, Kyoto 606-8507, Japan. 4. Department of Otolaryngology-Head and Neck Surgery, Kobe City Medical Center General Hospital, Minatojima-Minamimachi 2-1-1, Chuo-ku, Kobe 650-0047, Japan. 5. Department of Pathology, Kobe City Medical Center General Hospital, Minatojima-Minamimachi 2-1-1, Chuo-ku, Kobe 650-0047, Japan. 6. Department of Pathology, Kakogawa Central City Hospital, Honmachi 439, Kakogawa-cho, Kakogawa 675-8611, Japan. 7. Department of Neural Regeneration and Cell Communication, Mie University Graduate School of Medicine, Edobashi 2-174, Tsu 514-8507, Japan.
Abstract
OBJECTIVES: Confocal laser endomicroscopy (CLE) is a technology that enables microscopic visualization of lesions in real-time (optical biopsy) and has been successfully applied for clinical use in gastroenterology. Recently, it was also introduced for head and neck squamous cell carcinoma (HNSCC) diagnostics. We previously designed a self-made CLE, which can provide bichrome images, with topical contrast agents that are safe for use in patients. Herein, we report findings of a pilot study using our self-made CLE to image pairs of normal and cancerous tissues. This study aimed to characterize the features of HNSCC compared with normal mucosa and to establish a methodology of in vivo real-time optical biopsy of HNSCCs. METHODS: HNSCC tissues were acquired from 10 patients who underwent surgical resection. Dissected specimens were first evaluated for their auto-fluorescence spectral profiles with 473 nm laser excitation and further optical observation. While obtaining the image, auto-fluorescence spectrum and intensity of the reflectance fluorescent signals were measured in real-time by a spectrometer. Subsequently, acriflavine was applied to the specimen to fluorescently label the nuclei and observe the difference between normal and cancerous tissues with 473 nm laser excitation. Finally, double staining with acriflavine and edible Food Red No.106 was performed to observe both nuclei and the cytoplasm of normal and cancerous tissues at 473 nm and 561 nm laser excitation. RESULTS: Lower signals were detected from auto-fluorescence images of cancer tissues than normal tissues with 473 nm laser excitation. After acriflavine application, there was a clear difference between cancer and normal mucosa in the uniformity of nuclear size and shape. In normal mucosa, cells were arranged in an orderly manner, with each cell resembling a frog's egg. By contrast, in cancer tissues, the cell density was higher, and the cellular arrangement was less orderly. Using both acriflavine and Food Red No.106, images became more vivid, but more complicated because red dye staining of the cytoplasm emerged as fluorescence at different wavelengths. CONCLUSIONS: Real-time in vivo imaging using the newly developed CLE and conditions may be used to distinguish cancer tissue from normal mucosa without invasive biopsy.
OBJECTIVES: Confocal laser endomicroscopy (CLE) is a technology that enables microscopic visualization of lesions in real-time (optical biopsy) and has been successfully applied for clinical use in gastroenterology. Recently, it was also introduced for head and neck squamous cell carcinoma (HNSCC) diagnostics. We previously designed a self-made CLE, which can provide bichrome images, with topical contrast agents that are safe for use in patients. Herein, we report findings of a pilot study using our self-made CLE to image pairs of normal and cancerous tissues. This study aimed to characterize the features of HNSCC compared with normal mucosa and to establish a methodology of in vivo real-time optical biopsy of HNSCCs. METHODS:HNSCC tissues were acquired from 10 patients who underwent surgical resection. Dissected specimens were first evaluated for their auto-fluorescence spectral profiles with 473 nm laser excitation and further optical observation. While obtaining the image, auto-fluorescence spectrum and intensity of the reflectance fluorescent signals were measured in real-time by a spectrometer. Subsequently, acriflavine was applied to the specimen to fluorescently label the nuclei and observe the difference between normal and cancerous tissues with 473 nm laser excitation. Finally, double staining with acriflavine and edible Food Red No.106 was performed to observe both nuclei and the cytoplasm of normal and cancerous tissues at 473 nm and 561 nm laser excitation. RESULTS: Lower signals were detected from auto-fluorescence images of cancer tissues than normal tissues with 473 nm laser excitation. After acriflavine application, there was a clear difference between cancer and normal mucosa in the uniformity of nuclear size and shape. In normal mucosa, cells were arranged in an orderly manner, with each cell resembling a frog's egg. By contrast, in cancer tissues, the cell density was higher, and the cellular arrangement was less orderly. Using both acriflavine and Food Red No.106, images became more vivid, but more complicated because red dye staining of the cytoplasm emerged as fluorescence at different wavelengths. CONCLUSIONS: Real-time in vivo imaging using the newly developed CLE and conditions may be used to distinguish cancer tissue from normal mucosa without invasive biopsy.
Authors: Sneha Sethi; Xiangqun Ju; Richard M Logan; Paul Sambrook; Robert A McLaughlin; Lisa M Jamieson Journal: Int J Environ Res Public Health Date: 2021-11-25 Impact factor: 3.390
Authors: Matti Sievert; Konstantinos Mantsopoulos; Sarina K Mueller; Markus Eckstein; Robin Rupp; Marc Aubreville; Florian Stelzle; Nicolai Oetter; Andreas Maier; Heinrich Iro; Miguel Goncalves Journal: Acta Otorhinolaryngol Ital Date: 2022-02-07 Impact factor: 2.618