Fluorochrome‐dependent specific changes in spectral profiles using different compensation beads or primary cells in full spectrum cytometry Article Swipe
Yaroslava Shevchenko
,
Isabella Lurje
,
Frank Tacke
,
Linda Hammerich
·
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.1002/cyto.a.24836
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.1002/cyto.a.24836
Full spectrum flow cytometry is a powerful tool for immune monitoring on a single‐cell level and with currently available machines, panels of 40 or more markers per sample are possible. However, with an increased panel size, spectral unmixing issues arise, and appropriate single stain reference controls are required for accurate experimental results and to avoid unmixing errors. In contrast to conventional flow cytometry, full spectrum flow cytometry takes into account even minor differences in spectral signatures and requires the full spectrum of each fluorochrome to be identical in the reference control and the fully stained sample to ensure accurate and reliable results. In general, using the cells of interest is considered optimal, but certain markers may not be expressed at sufficient levels to generate a reliable positive control. In this case, compensation beads show some significant advantages as they bind a consistent amount of antibody independent of its specificity. In this study, we evaluated two types of manufactured compensation beads for use as reference controls for 30 of the most commonly used and commercially available fluorochromes in full spectrum cytometry and compared them to human and murine primary leukocytes. While most fluorochromes show the same spectral profile on beads and cells, we demonstrate that specific fluorochromes show a significantly different spectral profile depending on which type of compensation beads is used, and some fluorochromes should be used on cells exclusively. Here, we provide a list of important considerations when selecting optimal reference controls for full spectrum flow cytometry.
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Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1002/cyto.a.24836
- https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/cyto.a.24836
- OA Status
- hybrid
- Cited By
- 5
- References
- 16
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4393042616
All OpenAlex metadata
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- OpenAlex ID
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https://openalex.org/W4393042616Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1002/cyto.a.24836Digital Object Identifier
- Title
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Fluorochrome‐dependent specific changes in spectral profiles using different compensation beads or primary cells in full spectrum cytometryWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
-
2024Year of publication
- Publication date
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2024-03-21Full publication date if available
- Authors
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Yaroslava Shevchenko, Isabella Lurje, Frank Tacke, Linda HammerichList of authors in order
- Landing page
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https://doi.org/10.1002/cyto.a.24836Publisher landing page
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https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/cyto.a.24836Direct link to full text PDF
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YesWhether a free full text is available
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hybridOpen access status per OpenAlex
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https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/cyto.a.24836Direct OA link when available
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Flow cytometry, Cytometry, Stain, Compensation (psychology), Biological system, Computer science, Biology, Immunology, Staining, Genetics, Psychoanalysis, PsychologyTop concepts (fields/topics) attached by OpenAlex
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5Total citation count in OpenAlex
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2025: 4, 2024: 1Per-year citation counts (last 5 years)
- References (count)
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16Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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