Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.
Proteomics Program, Novo Nordisk Foundation Centre for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Methods Mol Biol. 2024;2817:97-113. doi: 10.1007/978-1-0716-3934-4_9.
Spatially resolved mass spectrometry-based proteomics at single-cell resolution promises to provide insights into biological heterogeneity. We describe a protocol based on multiplexed data-independent acquisition (mDIA) with dimethyl labeling to enhance proteome depth, accuracy, and throughput while minimizing costs. It enables high-quality proteome analysis of single isolated hepatocytes and utilizes liver zonation for single-cell proteomics benchmarking. This adaptable, modular protocol will promote the use of single-cell proteomics in spatial biology.
基于空间分辨质谱的蛋白质组学在单细胞分辨率上有望提供对生物异质性的深入了解。我们描述了一种基于多重数据非依赖采集（mDIA）和二甲化标记的方案，以提高蛋白质组的深度、准确性和通量，同时最小化成本。它能够实现单个分离肝细胞的高质量蛋白质组分析，并利用肝区带进行单细胞蛋白质组学基准测试。这种适应性强、模块化的方案将促进单细胞蛋白质组学在空间生物学中的应用。
