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Spatial Proteomics of Single Hepatocytes with Multiplexed Data-Independent Acquisition (mDIA)

  • Marvin Thielert,
  • Caroline A. M. Weiss,
  • Matthias Mann,
  • Florian A. Rosenberger

摘要

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.