Yun Dae Hee, Park Young-Gyun, Cho Jae Hun, Kamentsky Lee, Evans Nicholas B, DiNapoli Nicholas, Xie Katherine, Choi Seo Woo, Albanese Alexandre, Tian Yuxuan, Sohn Chang Ho, Zhang Qiangge, Kim Minyoung E, Swaney Justin, Guan Webster, Park Juhyuk, Drummond Gabi, Choi Heejin, Ruelas Luzdary, Feng Guoping, Chung Kwanghun
Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA.
Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA.
Nat Biotechnol. 2025 Jan 24. doi: 10.1038/s41587-024-02533-4.
Extending single-cell analysis to intact tissues while maintaining organ-scale spatial information poses a major challenge due to unequal chemical processing of densely packed cells. Here we introduce Continuous Redispersion of Volumetric Equilibrium (CuRVE) in nanoporous matrices, a framework to address this challenge. CuRVE ensures uniform processing of all cells in organ-scale tissues by perpetually maintaining dynamic equilibrium of the tissue's gradually shifting chemical environment. The tissue chemical reaction environment changes at a continuous, slow rate, allowing redispersion of unevenly distributed chemicals and preserving chemical equilibrium tissue wide at any given moment. We implemented CuRVE to immunologically label whole mouse and rat brains and marmoset and human tissue blocks within 1 day. We discovered highly variable regionalized reduction of parvalbumin immunoreactive cells in wild-type adult mice, a phenotype missed by the commonly used genetic labeling. We envision that our platform will advance volumetric single-cell processing and analysis, facilitating comprehensive single-cell level investigations within their spatial context in organ-scale tissues.
由于紧密堆积的细胞存在化学处理不均等问题，在保持器官尺度空间信息的同时将单细胞分析扩展到完整组织面临着重大挑战。在此，我们介绍了纳米多孔基质中的体积平衡连续再分散（CuRVE），这是一个应对这一挑战的框架。CuRVE通过永久维持组织逐渐变化的化学环境的动态平衡，确保对器官尺度组织中的所有细胞进行均匀处理。组织化学反应环境以连续、缓慢的速率变化，使得分布不均的化学物质得以再分散，并在任何给定时刻保持整个组织的化学平衡。我们应用CuRVE在1天内对整个小鼠和大鼠大脑以及狨猴和人类组织块进行了免疫标记。我们发现野生型成年小鼠中小清蛋白免疫反应性细胞存在高度可变的区域化减少，这是常用基因标记所遗漏的一种表型。我们设想我们的平台将推动体积单细胞处理和分析，促进在器官尺度组织的空间背景下进行全面的单细胞水平研究。
