Yau Chun Ngo, Hung Jacky Tin Shing, Campbell Robert A A, Wong Thomas Chun Yip, Huang Bei, Wong Ben Tin Yan, Chow Nick King Ngai, Zhang Lichun, Tsoi Eldric Pui Lam, Tan Yuqi, Li Joshua Jing Xi, Wing Yun Kwok, Lai Hei Ming
Department of Psychiatry, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong SAR, China.
Li Ka Shing Institute of Health Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong SAR, China.
Nat Commun. 2024 Dec 30;15(1):10888. doi: 10.1038/s41467-024-55248-0.
Biological systems are complex, encompassing intertwined spatial, molecular and functional features. However, methodological constraints limit the completeness of information that can be extracted. Here, we report the development of INSIHGT, a non-destructive, accessible three-dimensional (3D) spatial biology method utilizing superchaotropes and host-guest chemistry to achieve homogeneous, deep penetration of macromolecular probes up to centimeter scales, providing reliable semi-quantitative signals throughout the tissue volume. Diverse antigens, mRNAs, neurotransmitters, and post-translational modifications are well-preserved and simultaneously visualized. INSIHGT also allows multi-round, highly multiplexed 3D molecular probing and is compatible with downstream traditional histology and nucleic acid sequencing. With INSIHGT, we map undescribed podocyte-to-parietal epithelial cell microfilaments in mouse glomeruli and neurofilament-intensive inclusion bodies in the human cerebellum, and identify NPY-proximal cell types defined by spatial morpho-proteomics in mouse hypothalamus. We anticipate that INSIHGT can form the foundations for 3D spatial multi-omics technology development and holistic systems biology studies.
生物系统十分复杂，包含相互交织的空间、分子和功能特征。然而，方法上的限制阻碍了可提取信息的完整性。在此，我们报告了INSIHGT的开发，这是一种非破坏性的、易于操作的三维（3D）空间生物学方法，利用超级离液剂和主客体化学实现大分子探针在厘米尺度上的均匀、深度渗透，在整个组织体积内提供可靠的半定量信号。多种抗原、信使核糖核酸、神经递质和翻译后修饰都能得到良好保存并同时可视化。INSIHGT还允许进行多轮、高度多重的3D分子探测，并且与下游传统组织学和核酸测序兼容。借助INSIHGT，我们绘制了小鼠肾小球中未描述的足细胞到壁层上皮细胞的微丝以及人类小脑中神经丝密集的包涵体图谱，并通过空间形态蛋白质组学确定了小鼠下丘脑中与神经肽Y相邻的细胞类型。我们预计INSIHGT可为3D空间多组学技术发展和整体系统生物学研究奠定基础。
