ArticleAnalytical chemistry2020
Automated Coupling of Nanodroplet Sample Preparation with Liquid Chromatography-Mass Spectrometry for High-Throughput Single-Cell Proteomics.
Article in Analytical chemistry, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 85 papers.
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Who cites it
85 citing papers in PubMed, 162 citations in OpenAlex.
- Novel Cell-Type-Specific Drought-Responsive Proteins in Root Tips of Field-Grown Perennial Switchgrass.ACS omega · 2026Article
- Optimization of trap column properties and loading conditions for proteome profiling of single-cell-level sample inputs.Analytical and bioanalytical chemistry · 2026Article
- A 3D spatial proteomic map of the human pancreatic islet microenvironment.Research square · 2026Article
- Accelerated protein digestion by numbering-up nanochannels, followed by LC-MS analysis.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2026Article
- Integrating Spatial Proteogenomics in Cancer Research.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Review
- Evaluation and application of chemical decrosslinking in the context of histopathological spatial proteomics.bioRxiv : the preprint server for biology · 2026Article
- Modification of a Low-Cost Pipetting Robot for Nanoliter Liquid Handling and Autosampling for Liquid Chromatography-Mass Spectrometry.Journal of separation science · 2026Article
- Single-Islet Proteomics Maps Pseudo-Temporal Islet Immune Responses and Dysfunction in Stage 1 Type 1 Diabetes.bioRxiv : the preprint server for biology · 2025Article
- SLB-msSIM: A Spectral Library-Based Multiplex Segmented SIM Platform for Single-Cell Proteomic Analysis.Proteomics · 2025Article
- Imaging and spatially resolved mass spectrometry applications in nephrology.Nature reviews. Nephrology · 2025Review
- Oocyte mitochondria link maternal environment to offspring phenotype.bioRxiv : the preprint server for biology · 2025Article
- Delineation of Subcellular Molecular Heterogeneity in Single Cells via Ultralow-Flow-Rate Desorption Electrospray Ionization Mass Spectrometry Imaging.Analytical chemistry · 2025Article
- Increasing Proteome Coverage Through a Reduction in Analyte Complexity in Single-Cell Equivalent Samples.Journal of proteome research · 2025Article
- Cell Storage Conditions Impact Single-Cell Proteomic Landscapes.Journal of proteome research · 2025Article
- Digital Microfluidics for Sample Preparation in Low-Input Proteomics.Small methods · 2025Review
- Data acquisition approaches for single cell proteomics.Proteomics · 2025Review
- Innovative Advances in Droplet Microfluidics.Research (Washington, D.C.) · 2025Review
- Single-cell omics: experimental workflow, data analyses and applications.Science China. Life sciences · 2025Review
- Spatial Proteomics towards cellular Resolution.Expert review of proteomics · 2024Article
- Gradient-Elution Nanoflow Liquid Chromatography Without a Binary Pump: Smoothed Step Gradients Enable Reproducible, Sensitive, and Low-Cost Separations for Single-Cell Proteomics.Molecular & cellular proteomics : MCP · 2024Article
25 more citing papers are in PubMed but not listed here.
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Authors and funding
12 authors at 2 institutions in 1 country.
Funding
Abstract
Single-cell proteomics can provide critical biological insight into the cellular heterogeneity that is masked by bulk-scale analysis. We have developed a nanoPOTS (nanodroplet processing in one pot for trace samples) platform and demonstrated its broad applicability for single-cell proteomics. However, because of nanoliter-scale sample volumes, the nanoPOTS platform is not compatible with automated LC-MS systems, which significantly limits sample throughput and robustness. To address this challenge, we have developed a nanoPOTS autosampler allowing fully automated sample injection from nanowells to LC-MS systems. We also developed a sample drying, extraction, and loading workflow to enable reproducible and reliable sample injection. The sequential analysis of 20 samples containing 10 ng tryptic peptides demonstrated high reproducibility with correlation coefficients of >0.995 between any two samples. The nanoPOTS autosampler can provide analysis throughput of 9.6, 16, and 24 single cells per day using 120, 60, and 30 min LC gradients, respectively. As a demonstration for single-cell proteomics, the autosampler was first applied to profiling protein expression in single MCF10A cells using a label-free approach. At a throughput of 24 single cells per day, an average of 256 proteins was identified from each cell and the number was increased to 731 when the Match Between Runs algorithm of MaxQuant was used. Using a multiplexed isobaric labeling approach (TMT-11plex), ∼77 single cells could be analyzed per day. We analyzed 152 cells from three acute myeloid leukemia cell lines, resulting in a total of 2558 identified proteins with 1465 proteins quantifiable (70% valid values) across the 152 cells. These data showed quantitative single-cell proteomics can cluster cells to distinct groups and reveal functionally distinct differences.
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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.