Fluorescence microscopy of parathyroid and thyroid tissues for localization of autofluorescent substances using near-infrared wavelengths

IF 1.6 4区医学 Q2 OTORHINOLARYNGOLOGY

Auris Nasus Larynx Pub Date : 2025-02-01 DOI:10.1016/j.anl.2024.12.007

Yui Mizumoto-Teramura , Akira Leon Yoshikawa , Naoyuki Matsumoto , Yoko Murayama , Ken Akashi

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引用次数: 0

Abstract

Objective

The parathyroid gland emits autofluorescence with a peak at 822 nm when excited using near-infrared light at 785 nm; this observation of autofluorescence using a near-infrared detection device is useful for identifying the parathyroid gland during surgery. We aimed to clarify the localization of autofluorescent substances in parathyroid and thyroid tissues by observing them under a fluorescence microscope through filters that selectively pass specific near-infrared wavelengths.

Methods

Four cases of parathyroid and three cases of thyroid were examined under a fluorescence microscope. The frozen, formalin-fixed paraffin-embedded, and unfixed, unstained sections of parathyroid were observed through filters that selectively pass specific near-infrared wavelengths. Images were acquired at excitation 775 ± 50 nm and absorption 845 ± 55 nm in five randomly selected fields of view, avoiding tumor and inflammatory areas. Autofluorescence was measured as the ratio of fluorescent area to tissue area using hybrid cell counting.

Results

Autofluorescence was observed in all sections. In the parathyroid tissue, the frozen sections showed significantly more autofluorescence than the formalin-fixed paraffin-embedded sections, and in the thyroid tissue, although no significant difference was observed, the frozen sections showed more autofluorescence than the formalin-fixed paraffin-embedded sections. In addition, the single unfixed, unstained section showed stronger autofluorescence than the frozen sections, although no significant difference was found. The areas of autofluorescence in the parathyroid and thyroid tissues were thought to be the Golgi area and lipofuscin, respectively.

Conclusion

Fluorescence microscopy of parathyroid and thyroid tissues revealed the localization of autofluorescent substances in each tissue.

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使用近红外波长的甲状旁腺和甲状腺组织的荧光显微镜定位自身荧光物质。

目的：当使用波长为785 nm的近红外线激发甲状旁腺时，甲状旁腺会发出峰值为822 nm的自发荧光；使用近红外检测设备观察自发荧光有助于在手术中识别甲状旁腺。我们的目的是通过选择性通过特定近红外波长的滤光片在荧光显微镜下观察甲状旁腺和甲状腺组织中的自发荧光物质，从而明确它们的定位：方法：在荧光显微镜下观察4例甲状旁腺和3例甲状腺组织。通过选择性通过特定近红外波长的滤光片观察甲状旁腺冷冻、福尔马林固定、石蜡包埋和未固定、未染色的切片。在随机选择的五个视野中，以激发波长 775 ± 50 nm 和吸收波长 845 ± 55 nm 获取图像，避开肿瘤和炎症区域。采用杂交细胞计数法测量自发荧光，即荧光面积与组织面积之比：所有切片都观察到了自发荧光。在甲状旁腺组织中，冷冻切片显示的自发荧光明显多于福尔马林固定的石蜡包埋切片；在甲状腺组织中，虽然没有观察到显著差异，但冷冻切片显示的自发荧光多于福尔马林固定的石蜡包埋切片。此外，未固定、未染色的单个切片比冷冻切片显示出更强的自发荧光，但未发现明显差异。甲状旁腺和甲状腺组织的自发荧光区域被认为分别是高尔基体区和脂褐素：甲状旁腺和甲状腺组织的荧光显微镜检查显示了自发荧光物质在各组织中的定位。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Auris Nasus Larynx 医学-耳鼻喉科学

CiteScore

3.40

自引率

5.90%

发文量

169

审稿时长

30 days

期刊介绍： The international journal Auris Nasus Larynx provides the opportunity for rapid, carefully reviewed publications concerning the fundamental and clinical aspects of otorhinolaryngology and related fields. This includes otology, neurotology, bronchoesophagology, laryngology, rhinology, allergology, head and neck medicine and oncologic surgery, maxillofacial and plastic surgery, audiology, speech science. Original papers, short communications and original case reports can be submitted. Reviews on recent developments are invited regularly and Letters to the Editor commenting on papers or any aspect of Auris Nasus Larynx are welcomed. Founded in 1973 and previously published by the Society for Promotion of International Otorhinolaryngology, the journal is now the official English-language journal of the Oto-Rhino-Laryngological Society of Japan, Inc. The aim of its new international Editorial Board is to make Auris Nasus Larynx an international forum for high quality research and clinical sciences.