High-Throughput Single-Cell Proteomic Analysis of Organ-Derived Heterogeneous Cell Populations by Nanoflow Dual-Trap Single-Column Liquid Chromatography.
Publication Year:
2023
PubMed ID:
37289937
Funding Grants:
Public Summary:
This study introduces a new method called nanoDTSC, which is a fast and efficient way to analyze proteins in individual cells using mass spectrometry. This technique can process 96 single cells per day, identifying over 1,000 proteins in heart muscle cells and mixed cell types from the aorta. Using standard lab equipment, nanoDTSC allows researchers to quickly study rare cell types within complex tissues without needing special labels, making it easier to understand the detailed protein makeup of individual cells.
Scientific Abstract:
Identification and proteomic characterization of rare cell types within complex organ-derived cell mixtures is best accomplished by label-free quantitative mass spectrometry. High throughput is required to rapidly survey hundreds to thousands of individual cells to adequately represent rare populations. Here we present parallelized nanoflow dual-trap single-column liquid chromatography (nanoDTSC) operating at 15 min of total run time per cell with peptides quantified over 11.5 min using standard commercial components, thus offering an accessible and efficient LC solution to analyze 96 single cells per day. At this throughput, nanoDTSC quantified over 1000 proteins in individual cardiomyocytes and heterogeneous populations of single cells from the aorta.