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

摘要 先天性巨结肠病是一种先天性缺陷，影响估计1：5000活产婴儿。它 其特征在于远端直肠中缺少神经节细胞（神经元）， 邻近近端肠。先天性巨结肠的诊断依赖于主观的 病理学家对来自抽吸直肠活检的多达100个组织切片的解释，以证明 缺乏神经节细胞和存在肥大神经。由于劳动密集型 和耗时的性质，目前的诊断过程，有必要更有效地 综合诊断先天性巨结肠。我们的团队率先使用3D技术 开放式光片（OTLS）显微镜，它能够快速，高通量成像的大 临床样本。结合尖端的机器学习技术，我们假设 3D OTLS显微镜可以提供更准确和一致的神经节评估， 疑似直肠癌患者抽吸直肠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检验这一假设 和机器学习分析管道，将在100个直肠活检标本上进行试点 新鲜的切除标本。

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.