ArticleAnalytical and bioanalytical chemistry2019
Benchtop-compatible sample processing workflow for proteome profiling of < 100 mammalian cells.
Article in Analytical and bioanalytical chemistry, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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Who cites it
36 citing papers in PubMed, 57 citations in OpenAlex.
- Single-Islet Proteomics Maps Pseudo-Temporal Islet Immune Responses and Dysfunction in Stage 1 Type 1 Diabetes.bioRxiv : the preprint server for biology · 2025Article
- Activity-targeted metaproteomics uncovers rare syntrophic bacteria central to anaerobic community metabolism.Nature microbiology · 2025Article
- The $10 proteome: low-cost, deep and quantitative proteome profiling of limited sample amounts using the Orbitrap Astral and timsTOF Ultra 2 mass spectrometers.bioRxiv : the preprint server for biology · 2025Article
- Alterations of the skeletal muscle nuclear proteome after acute exercise reveals a posttranscriptional influence.American journal of physiology. Cell physiology · 2025Article
- Coupling Microdroplet-Based Sample Preparation, Multiplexed Isobaric Labeling, and Nanoflow Peptide Fractionation for Deep Proteome Profiling of the Tissue Microenvironment.Analytical chemistry · 2024Article
- Instrumentation at the Leading Edge of Proteomics.Analytical chemistry · 2024Review
- Dilute to Enrich for Deeper Proteomics: A Yolk-Depleted Carrier for Limited Populations of Embryonic (Frog) Cells.Journal of proteome research · 2024Article
- Infrared Laser Ablation and Capture of Biological Tissue.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Efficient and Sensitive Sample Preparation, Separations, and Data Acquisition for Label-Free Single-Cell Proteomics.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Easy and Accessible Workflow for Label-Free Single-Cell Proteomics.Journal of the American Society for Mass Spectrometry · 2023Article
- Rapid, One-Step Sample Processing for Label-Free Single-Cell Proteomics.Journal of the American Society for Mass Spectrometry · 2023Article
- Proteome Mapping of the Human Pancreatic Islet Microenvironment Reveals Endocrine-Exocrine Signaling Sphere of Influence.Molecular & cellular proteomics : MCP · 2023Article
- Unlocking the potential of microfluidics in mass spectrometry-based immunopeptidomics for tumor antigen discovery.Cell reports methods · 2023Review
- All-in-One digital microfluidics pipeline for proteomic sample preparation and analysis.Chemical science · 2023Article
- A streamlined tandem tip-based workflow for sensitive nanoscale phosphoproteomics.Communications biology · 2023Article
- Untangling Alzheimer's disease with spatial multi-omics: a brief review.Frontiers in aging neuroscience · 2023Review
- Enhanced Spatial Mapping of Histone Proteoforms in Human Kidney Through MALDI-MSI by High-Field UHMR-Orbitrap Detection.Analytical chemistry · 2022Article
- In-Depth Mass Spectrometry-Based Proteomics of Formalin-Fixed, Paraffin-Embedded Tissues with a Spatial Resolution of 50-200 μm.Journal of proteome research · 2022Article
- Interfacing microfluidics with information-rich detection systems for cells, bioparticles, and molecules.Analytical and bioanalytical chemistry · 2022Review
- Evaluation of Fast and Sensitive Proteome Profiling of FF and FFPE Kidney Patient Tissues.Molecules (Basel, Switzerland) · 2022Article
Corrections and comments
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Authors and funding
10 authors at 3 institutions in 1 country.
Funding
Abstract
Extending proteomics to smaller samples can enable the mapping of protein expression across tissues with high spatial resolution and can reveal sub-group heterogeneity. However, despite the continually improving sensitivity of LC-MS instrumentation, in-depth profiling of samples containing low-nanogram amounts of protein has remained challenging due to analyte losses incurred during preparation and analysis. To address this, we recently developed nanodroplet processing in one pot for trace samples (nanoPOTS), a robotic/microfluidic platform that generates ready-to-analyze peptides from cellular material in ~200 nL droplets with greatly reduced sample losses. In combination with ultrasensitive LC-MS, nanoPOTS has enabled >3000 proteins to be confidently identified from as few as 10 cultured human cells and ~700 proteins from single cells. However, the nanoPOTS platform requires a highly skilled operator and a costly in-house-built robotic nanopipetting instrument. In this work, we sought to evaluate the extent to which the benefits of nanodroplet processing could be preserved when upscaling reagent dispensing volumes by a factor of 10 to those addressable by commercial micropipette. We characterized the resulting platform, termed microdroplet processing in one pot for trace samples (μPOTS), for the analysis of as few as ~25 cultured HeLa cells (4 ng total protein) or 50 μm square mouse liver tissue thin sections and found that ~1800 and ~1200 unique proteins were respectively identified with high reproducibility. The reduced equipment requirements should facilitate broad dissemination of nanoproteomics workflows by obviating the need for a capital-intensive custom liquid handling system.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.