Abstract

Single-cell tandem MS has enabled analyzing hundreds of single cells per day and quantifying thousands of proteins across the cells. The broad dissemination of these capabilities can empower the dissection of pathophysiological mechanisms in heterogeneous tissues. Key requirements for achieving this goal include robust protocols performed on widely accessible hardware, robust quality controls, community standards, and automated data analysis pipelines that can pinpoint analytical problems and facilitate their timely resolution. Toward meeting these requirements, this perspective outlines both existing resources and outstanding opportunities, such as parallelization, for catalyzing the wide dissemination of quantitative single-cell proteomics analysis that can be scaled up to tens of thousands of single cells. Indeed, simultaneous parallelization of the analysis of peptides and single cells is a promising approach for multiplicative increase in the speed of performing deep and quantitative single-cell proteomics. The community is ready to begin a virtuous cycle of increased adoption fueling the development of more technology and resources for single-cell proteomics that in turn drive broader adoption, scientific discoveries, and clinical applications.

Highlights

  • Protocols for single-cell proteomics by MS are ready for broad adoption

  • Sample preparation throughput increased with the introduction of automated multiwell-plate methods, such as minimal ProteOmic sample Preparation [21, 39, 40] and automated preparation in one pot for trace samples [41]

  • Increasing appreciation for the need to perform single-cell protein analysis has propelled the field of single-cell proteomics by MS, resulting in methods that allow quantifying over a thousand proteins per cell while analyzing hundreds of single cells per day

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Summary

Graphical Abstract

In Brief Single-cell proteomics will drive the wave of single-cell biology. This requires broad adoption of existing methods, the application of rigorous quality control standards, and the continuous advancement of the technology. The advancement will be driven by numerous innovations, including highly parallelized analysis, and will increase the throughput, quantitative accuracy, and the accessibility of the single-cell proteomics

Nikolai Slavov*
INCREASING ROBUSTNESS AND ACCESSIBILITY
Pipelines for data analysis and interpretation
HIGHLY PARALLEL SAMPLE PREPARATION
PARALLEL ANALYSIS OF BOTH PEPTIDES AND SINGLE CELLS
CONCLUSION
Full Text
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