3D diagnosis of Hirschsprung disease using open-top light-sheet microscopy

摘要 先天性巨结肠症是一种先天性缺陷，估计有1：5000的活产婴儿受到影响。它 其特征是直肠远端无神经节细胞(神经元)和可变长度的 邻近的近端肠道。先天性巨结肠的诊断依赖于主观判断 病理学家解释多达100个组织切片从吸引直肠活检，以证明 神经节细胞缺失，神经肥大。由于劳动密集型 而当前诊断过程的耗时性质，需要更有效地 全面诊断先天性巨结肠。我们的团队已经率先使用了3D 开顶式光片(OTLS)显微镜，支持快速、高通量成像 临床样本。结合尖端的机器学习技术，我们假设 3D OTLS显微镜可以为神经节提供更准确和一致的评估 疑似直肠抽吸活检中的细胞和肥大神经 先天性巨结肠。我们将通过开发一种多重染色方案来验证这一假设 和机器学习分析流水线，将在100个直肠活检样本上进行试点 从新鲜的切除标本中提取。

Abstract Hirschsprung disease is a congenital defect that affects an estimated 1:5000 liveborn infants. It is characterized by absent ganglion cells (neurons) in the distal rectum and a variable length of contiguous proximal bowel. The diagnosis of Hirschsprung disease relies upon subjective pathologist interpretation of up to 100 tissue sections from suction rectal biopsies to prove the absence of ganglion cells and the presence of hypertrophic nerves. Due to the labor-intensive and time-consuming nature of the current diagnostic process, there is a need to more efficiently and comprehensively diagnose Hirschsprung disease. Our team has pioneered the use of 3D open-top light-sheet (OTLS) microscopy, which enables rapid, high-throughput imaging of large clinical samples. In combination with cutting-edge machine learning techniques, we hypothesize that 3D OTLS microscopy can provide more accurate and consistent assessment of ganglion cells and hypertrophic nerves in suction rectal biopsies from patients with suspected Hirschsprung disease. We will test this hypothesis by developing a multiplex staining protocol and machine learning analysis pipeline, which will be piloted on 100 rectal biopsy specimens from fresh resection specimens.