ArticleNature protocols2019
Development, application and computational analysis of high-dimensional fluorescent antibody panels for single-cell flow cytometry.
Article in Nature protocols, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 104 papers.
What it found
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Who cites it
104 citing papers in PubMed.
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- Protocol to annotate dendritic cell maturation types in vivo making use of lipid nanoparticle-based approaches.STAR protocols · 2026Article
- Potent antitumor activity through dual targeting of PD-L1 and TGF-β pathways in the glioma tumor microenvironment.Journal for immunotherapy of cancer · 2026Article
- Multi-Scale Transcriptomics Redefining the Tumor Immune Microenvironment.Biotech (Basel (Switzerland)) · 2026Review
- Human NKT10 cells are enriched in cord-derived invariant natural killer T cells and mediate immune-regulation in a xenogeneic graft-versus-host disease model.Frontiers in immunology · 2026Article
- HIV-Exposed Seronegative Female Sex Workers Show Different Cellular Immune Profiles Across the Menstrual Cycle.American journal of reproductive immunology (New York, N.Y. : 1989) · 2025Article
- Antibody-Based Multiplex Image Analysis: Standard Analytical Workflows and Artificial Intelligence Tools for Pathologists.Laboratory investigation; a journal of technical methods and pathology · 2025Review
- cytoGPNet: Enhancing clinical outcome prediction accuracy using longitudinal cytometry data in small cohort studies.Patterns (New York, N.Y.) · 2025Article
- Comparative Analysis of Immune Cell Populations From Two Sampling Techniques of Human Term Decidua Utilizing High-Parameter Full-Spectrum Flow Cytometry.American journal of reproductive immunology (New York, N.Y. : 1989) · 2025Article
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- Aspirin prevents metastasis by limiting platelet TXANature · 2025Article
- Single-cell omics technologies - Fundamentals on how to create single-cell looking glasses for reproductive health.American journal of obstetrics and gynecology · 2025Review
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- Tracking immune dysregulation in COVID-19: lymphocyte dynamics from hospitalization to recovery.Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologica · 2025Article
- Touch as emotion regulation.Frontiers in psychology · 2025Review
- Impact of chemotherapy on humoral and cellular immune responses to COVID-19 vaccination in patients with solid tumors.Frontiers in immunology · 2025Article
44 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The interrogation of single cells is revolutionizing biology, especially our understanding of the immune system. Flow cytometry is still one of the most versatile and high-throughput approaches for single-cell analysis, and its capability has been recently extended to detect up to 28 colors, thus approaching the utility of cytometry by time of flight (CyTOF). However, flow cytometry suffers from autofluorescence and spreading error (SE) generated by errors in the measurement of photons mainly at red and far-red wavelengths, which limit barcoding and the detection of dim markers. Consequently, development of 28-color fluorescent antibody panels for flow cytometry is laborious and time consuming. Here, we describe the steps that are required to successfully achieve 28-color measurement capability. To do this, we provide a reference map of the fluorescence spreading errors in the 28-color space to simplify panel design and predict the success of fluorescent antibody combinations. Finally, we provide detailed instructions for the computational analysis of such complex data by existing, popular algorithms (PhenoGraph and FlowSOM). We exemplify our approach by designing a high-dimensional panel to characterize the immune system, but we anticipate that our approach can be used to design any high-dimensional flow cytometry panel of choice. The full protocol takes a few days to complete, depending on the time spent on panel design and data analysis.
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Registered trials
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