OBJECTIVE In living tissue, it has been difficult to make microscopic-level observations without damaging the tissue. SUMMARY BACKGROUND DATA We have invented a novel intravital fluorescent observation method (IFOM) for real-time tissue observation, combining multi-photon laser scanning microscopy (MPLSM) with curcumin vital staining (CVS-IFOM). The aim of this study was to use CVS-IFOM to analyze the enteric nervous system (ENS) in mice and human patients with hypoganglionosis and Hirschsprung disease. METHODS In an initial viability study, we compared live ENS images from non-fluorescent C57BL6 mice stained with curcumin (n=5) and GFP mice (n=5) using MPLSM. We then explored CVS-IFOM for the live examination of resected colon tissues from one hypoganglionosis and three Hirschsprung disease patients. RESULTS In the viability study, detailed ENS histological features were only observed in the curcumin-stained mice. In the hypoganglionosis patient, CVS-IFOM provided ENS details that were not visualized under H&E staining or calretinin immunohistochemistry, allowing the analysis of ENS size, neural bundle number, and neural cell number per plexus. In Hirschsprung disease patients, CVS-IFOM showed a gradual hypoplastic change in the ENS from the oral wedge to the anal wedge, detecting disproportionate changes in the ENS within the same intestinal level, supporting a circumferentially uneven distribution of the intestinal ENS. CONCLUSION CVS-IFOM may be supportive for intraoperative pathological diagnosis during surgeries in Hirschsprung disease.

中文翻译：

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使用姜黄素辅助多光子激光成像对先天性巨结肠进行肠道神经网络的动态病理学。


目的 在活体组织中，很难在不损伤组织的情况下进行微观观察。摘要背景数据我们发明了一种用于实时组织观察的新型活体荧光观察方法（IFOM），该方法将多光子激光扫描显微镜（MPLSM）与姜黄素活体染色（CVS-IFOM）相结合。本研究的目的是使用 CVS-IFOM 分析患有神经节减退症和先天性巨结肠的小鼠和人类患者的肠神经系统 (ENS)。方法 在最初的活力研究中，我们使用 MPLSM 比较了姜黄素染色的非荧光 C57BL6 小鼠 (n=5) 和 GFP 小鼠 (n=5) 的实时 ENS 图像。然后，我们探索了 CVS-IFOM 对一名神经节减退症和三名先天性巨结肠患者切除的结肠组织进行实时检查。结果 在活力研究中，仅在姜黄素染色的小鼠中观察到详细的 ENS 组织学特征。在神经节减退症患者中，CVS-IFOM 提供了 H&E 染色或钙视网膜蛋白免疫组织化学下未显示的 ENS 详细信息，从而可以分析 ENS 大小、神经束数量和每个神经丛的神经细胞数量。在先天性巨结肠症患者中，CVS-IFOM 显示 ENS 从口腔楔形到肛门楔形逐渐发育不全，检测到同一肠道水平内 ENS 的不成比例的变化，支持肠道 ENS 的周向不均匀分布。结论 CVS-IFOM可能有助于先天性巨结肠手术中的术中病理诊断。