{
  "abstract": "Advances in spectral cytometry instrumentation and fluorescent reagents have led to the possibility of ultra-high-parameter panels exceeding 50 colors. However, panel size is limited in practice by unmixing-dependent spreading (UDS), a phenomenon which leads to a progressive deterioration of unmixed signal-to-noise ratios in panels that contain fluorochrome combinations with significant spectral overlap. So far, choosing spectrally compatible sets of fluorochromes that avoid UDS has been a complex and labor-intensive task involving substantial trial-and-error experimentation. Here, we provide a detailed explanation of UDS and practical strategies for handling UDS in large spectral panels. We describe the empirical hallmarks of UDS, demonstrate how to quantify its impact, and dissect its underlying mathematical cause in terms of spectral collinearity. We present practical tools derived from the regression literature that can be used to select optimal combinations of fluorochromes in a platform-agnostic fashion based on publicly available reference data, providing a general tool for spectral panel design.",
  "abstract_shown": true,
  "affiliations": [
    "Advanced Technology Group Waters Biosciences (formerly BD Biosciences) Milpitas California USA",
    "Department of Immunology University of Washington Seattle Washington USA",
    "Vaccine and Infectious Disease Division Fred Hutchinson Cancer Center Seattle Washington USA",
    "Flow Cytometry Core Facility ETH Zurich Zurich Switzerland"
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 3,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Peter L. Mage",
      "orcid": "0000-0002-3930-2343"
    },
    {
      "affs": [
        1,
        2
      ],
      "name": "A Konecny",
      "orcid": "0000-0002-3492-1006"
    },
    {
      "affs": [
        3
      ],
      "name": "Florian Mair",
      "orcid": "0000-0001-6732-5449"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/77gg3ap8w1m4h.bib",
  "cited_by": 14,
  "doi": "10.1002/cyto.a.70044",
  "fulltext_url": null,
  "id": "77gg3ap8w1m4h",
  "issns": [
    "1552-4922",
    "1552-4930"
  ],
  "landing_url": "https://doi.org/10.1002/cyto.a.70044",
  "language": "en",
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1304|Advanced Fluorescence Microscopy Techniques"
    ],
    [
      "openalex",
      "1304|Cell Image Analysis Techniques"
    ]
  ],
  "openalex": "W7164682476",
  "pdf_url": null,
  "pmcid": "PMC12139922",
  "pmid": "40475599",
  "published": "2026-06-01",
  "publisher": "Wiley",
  "ris": "https://research.draconitic.com/paper/77gg3ap8w1m4h.ris",
  "sources": [
    {
      "first_seen": "2026-11-04T13:21:05Z",
      "native_id": "W7164682476",
      "source": "openalex",
      "url": "https://doi.org/10.1002/cyto.a.70044"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1304|Advanced Fluorescence Microscopy Techniques",
      "subfield": "biophysics"
    }
  ],
  "title": "Measurement and Prediction of Unmixing‐Dependent Spreading due to Collinearity in Spectral Flow Cytometry",
  "title_text": "Measurement and Prediction of Unmixing‐Dependent Spreading due to Collinearity in Spectral Flow Cytometry",
  "type": "peer-reviewed",
  "updated_at": "2026-11-09T13:21:05Z",
  "url": "https://research.draconitic.com/paper/77gg3ap8w1m4h",
  "venue": "Cytometry Part A",
  "versions": []
}