高维成像的出现为以前依赖组织病理学定义的疾病的分子诊断细胞提供了新的机会。结节性硬化症（TSC）就是一个例子，这是一种以良性肿瘤全身生长为特征的发育障碍。在 TSC 患者切除的脑组织中，检测到异常增大的球囊细胞 (BC) 是这种疾病的特征。尽管神经病理学家可以鉴定 BC，但人们对这些细胞的蛋白质标记物的特异性和广泛适用性知之甚少，这使得在这种疾病的实验模型中鉴定的拟议 BC 的分类变得复杂。在这里，我们报告了定制机器学习管道（BAlloon IDENtifier；BAIDEN）的开发，该管道经过训练，可以使用与高维细胞计数兼容的组织学染色前瞻性地识别组织切片中的BC。该方法与定制的 36 抗体面板和成像质量流式细胞术 (IMC) 相结合，以探索先前提出的多种 BC 标记蛋白的表达，并开发 TSC 患者多个组织样本中保守的 BC 特征的描述符。在这里，我们提出了一个模块化工作流程，包括 BAIDEN、定制抗体面板、对照样品微阵列和分析管道（开源和内部），并应用此工作流程来了解 BC 的丰度、结构和信号活动，例如高维成像如何应用于人体组织的示例。© 2024 作者。细胞计数 B 部分：临床细胞计数由 Wiley periodicals LLC 代表国际临床细胞计数协会出版。

The advent of high-dimensional imaging offers new opportunities to molecularly characterize diagnostic cells in disorders that have previously relied on histopathological definitions. One example case is found in tuberous sclerosis complex (TSC), a developmental disorder characterized by systemic growth of benign tumors. Within resected brain tissues from patients with TSC, detection of abnormally enlarged balloon cells (BCs) is pathognomonic for this disorder. Though BCs can be identified by an expert neuropathologist, little is known about the specificity and broad applicability of protein markers for these cells, complicating classification of proposed BCs identified in experimental models of this disorder. Here, we report the development of a customized machine learning pipeline (BAlloon IDENtifier; BAIDEN) that was trained to prospectively identify BCs in tissue sections using a histological stain compatible with high-dimensional cytometry. This approach was coupled to a custom 36-antibody panel and imaging mass cytometry (IMC) to explore the expression of multiple previously proposed BC marker proteins and develop a descriptor of BC features conserved across multiple tissue samples from patients with TSC. Here, we present a modular workflow encompassing BAIDEN, a custom antibody panel, a control sample microarray, and analysis pipelines-both open-source and in-house-and apply this workflow to understand the abundance, structure, and signaling activity of BCs as an example case of how high-dimensional imaging can be applied within human tissues.© 2024 The Author(s). Cytometry Part B: Clinical Cytometry published by Wiley Periodicals LLC on behalf of International Clinical Cytometry Society.