Abstract:
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.
摘要:
单细胞分辨率的基于空间分辨质谱的蛋白质组学有望提供对生物异质性的见解。我们描述了一种基于复用数据无关采集(mDIA)与二甲基标记的协议,以增强蛋白质组深度,准确度,和吞吐量,同时降低成本。它可以对单个分离的肝细胞进行高质量的蛋白质组分析,并利用肝脏分区进行单细胞蛋白质组学基准测试。这种适应性,模块化协议将促进单细胞蛋白质组学在空间生物学中的应用。