biostudies-literatureUnknown17(10)Cai XHigh-throughput lysis and proteolytic digestion of biopsy-level tissue specimens is a major bottleneck for clinical proteomics. Here we describe a detailed protocol of pressure cycling technology (PCT)-assisted sample preparation for proteomic analysis of biopsy tissues. A piece of fresh frozen or formalin-fixed paraffin-embedded tissue weighing ~0.1-2 mg is placed in a 150 μL pressure-resistant tube called a PCT-MicroTube with proper lysis buffer. After closing with a PCT-MicroPestle, a batch of 16 PCT-MicroTubes are placed in a Barocycler, which imposes oscillating pressure to the samples from one atmosphere to up to ~3,000 times atmospheric pressure. The pressure cycling schemes are optimized for tissue lysis and protein digestion, and can be programmed in the Barocycler to allow reprodNature protocols2307-2325https://www.ebi.ac.uk/biostudies/studies/S-EPMC9362583biostudies-literatureHigh-throughput proteomic sample preparation using pressure cycling technology.PMC9362583Xu LYi XZhu YWu CYue LGuo TGao HXue ZChen CYu JLiu WGe WSun RQian LCai XfalseHigh-throughput proteomic sample preparation using pressure cycling technology.High-throughput lysis and proteolytic digestion of biopsy-level tissue specimens is a major bottleneck for clinical proteomics. Here we describe a detailed protocol of pressure cycling technology (PCT)-assisted sample preparation for proteomic analysis of biopsy tissues. A piece of fresh frozen or formalin-fixed paraffin-embedded tissue weighing ~0.1-2 mg is placed in a 150 μL pressure-resistant tube called a PCT-MicroTube with proper lysis buffer. After closing with a PCT-MicroPestle, a batch of 16 PCT-MicroTubes are placed in a Barocycler, which imposes oscillating pressure to the samples from one atmosphere to up to ~3,000 times atmospheric pressure. The pressure cycling schemes are optimized for tissue lysis and protein digestion, and can be programmed in the Barocycler to allow reprod2022-01-01T00:00:00Z2022 Oct2025-04-05T23:57:35.254Z2025-04-05T23:57:35.254ZS-EPMC93625833593177810.1038/s41596-022-00727-1