Normalization of mass cytometry data with bead standards
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Résumé
Mass cytometry uses atomic mass spectrometry combined with isotopically pure reporter elements to currently measure as many as 40 parameters per single cell. As with any quantitative technology, there is a fundamental need for quality assurance and normalization protocols. In the case of mass cytometry, the signal variation over time due to changes in instrument performance combined with intervals between scheduled maintenance must be accounted for and then normalized. Here, samples were mixed with polystyrene beads embedded with metal lanthanides, allowing monitoring of mass cytometry instrument performance over multiple days of data acquisition. The protocol described here includes simultaneous measurements of beads and cells on the mass cytometer, subsequent extraction of the bead-based signature, and the application of an algorithm enabling correction of both short- and long-term signal fluctuations. The variation in the intensity of the beads that remains after normalization may also be used to determine data quality. Application of the algorithm to a one-month longitudinal analysis of a human peripheral blood sample reduced the range of median signal fluctuation from 4.9-fold to 1.3-fold.
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La notice
- Revue
- Cytometry Part A
- Thématique
- Single-cell and spatial transcriptomics
- Domaine
- Biochemistry, Genetics and Molecular Biology
- Établissements canadiens
- —
- Organismes subventionnaires
- National Institute of Allergy and Infectious DiseasesU.S. Public Health ServiceNational Cancer InstituteNational Institutes of HealthNational Eye InstituteUniversity of TorontoDamon Runyon Cancer Research Foundation
- Mots-clés
- Mass cytometryNormalization (sociology)CytometryReproducibilityQuality assuranceMass spectrometryFlow cytometryComputer scienceBiomedical engineeringMaterials scienceChemistryAnalytical Chemistry (journal)Biological systemChromatographyMedicineBiologyPathologyExternal quality assessmentMolecular biology
- Résumé présent dans OpenAlex
- oui