类有机物
细胞生物学
细胞
质量细胞仪
电池类型
生物
流式细胞术
细胞信号
单细胞分析
信号转导
表型
分子生物学
生物化学
基因
作者
Jahangir Sufi,Xiao Qin,Ferran Cardoso Rodriguez,Yong Jia Bu,Petra Vlckova,María Ramos Zapatero,Mark Nitz,Christopher J. Tape
出处
期刊:Nature Protocols
[Springer Nature]
日期:2021-09-08
卷期号:16 (10): 4897-4918
被引量:21
标识
DOI:10.1038/s41596-021-00603-4
摘要
Organoids are biomimetic tissue models comprising multiple cell types and cell states. Post-translational modification (PTM) signaling networks control cellular phenotypes and are frequently dysregulated in diseases such as cancer. Although signaling networks vary across cell types, there are limited techniques to study cell type-specific PTMs in heterocellular organoids. Here, we present a multiplexed mass cytometry (MC) protocol for single-cell analysis of PTM signaling and cell states in organoids and organoids co-cultured with fibroblasts and leukocytes. We describe how thiol-reactive organoid barcoding in situ (TOBis) enables 35-plex and 126-plex single-cell comparison of organoid cultures and provide a cytometry by time of flight (CyTOF) signaling analysis pipeline (CyGNAL) for computing cell type-specific PTM signaling networks. The TOBis MC protocol takes ~3 d from organoid fixation to data acquisition and can generate single-cell data for >40 antibodies from millions of cells across 126 organoid cultures in a single MC run.
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