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Module HEME-28 · Version 1.0

Flow Cytometry Fundamentals and Gating

Learn how a flow cytometer records single-cell events and how a reproducible gate hierarchy, controls, and stated denominator support a bounded hematology result.

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Who this module is for

Medical laboratory scientists and trainees with cell biology, antibody, and HEME-01 prerequisites who need a foundation for reviewing clinical hematology flow-cytometry data.

Learning objectives

  • Describe hydrodynamic focusing, scatter, fluorescence, and per-event collection.
  • Apply a reproducible sequence of debris, singlet, viability, lineage, and population gates.
  • Recognize spillover, autofluorescence, nonspecific binding, low event counts, and specimen-quality limits before interpreting a population.

How completion works

Mark every required section done and answer every knowledge check in those sections correctly. The final save completes the module automatically.

Sources

11 sources

  1. 1. 42 CFR §493.1253, Establishment and verification of performance specifications. Federal regulation. https://www.ecfr.gov/current/title-42/chapter-IV/subchapter-G/part-493/subpart-K/section-493.1253. Accessed 2026-08-16.

    Source note · federal regulation

  2. 2. 42 CFR §493.1256, Control procedures. Federal regulation. https://www.ecfr.gov/current/title-42/chapter-IV/subchapter-G/part-493/subpart-K/section-493.1256. Accessed 2026-08-16.

    Source note · federal regulation

  3. 3. Clinical and Laboratory Standards Institute. H43-A2: Clinical Flow Cytometric Analysis of Neoplastic Hematolymphoid Cells; Approved Guideline, 2nd ed. 2007, reaffirmed 2017. Consensus standard. https://clsi.org/shop/standards/h43/. Accessed 2026-08-16.

    Source note · consensus standard

  4. 4. Clinical and Laboratory Standards Institute. H62Ed1E: Validation of Assays Performed by Flow Cytometry, 1st ed. 2021. Consensus standard. https://clsi.org/shop/standards/h62/. Accessed 2026-08-16.

    Source note · consensus standard

  5. 5. Li W. Flow Cytometry in the Diagnosis of Leukemias. In: Li W, ed. Leukemia. Exon Publications; 2022. NCBI Bookshelf review. https://www.ncbi.nlm.nih.gov/books/NBK586209/. Accessed 2026-08-16.

    Source note · peer-reviewed literature

  6. 6. Cossarizza A, et al. Guidelines for the use of flow cytometry and cell sorting in immunological studies, third edition. European Journal of Immunology. 2021;51:2708-3145. Peer-reviewed consensus guideline. https://pmc.ncbi.nlm.nih.gov/articles/PMC11115438/. Accessed 2026-08-16.

    Source note · consensus standard

  7. 7. Cossarizza A, et al. Guidelines for the use of flow cytometry and cell sorting in immunological studies, third edition, data analysis sections. European Journal of Immunology. 2021;51:2708-3145. Peer-reviewed consensus guideline. https://pmc.ncbi.nlm.nih.gov/articles/PMC9165548/. Accessed 2026-08-16.

    Source note · consensus standard

  8. 8. Craig FE, Foon KA. Clinical flow-cytometric testing in chronic lymphocytic leukemia. Archives of Pathology & Laboratory Medicine. 2008;132:9-18. Peer-reviewed clinical review. https://pmc.ncbi.nlm.nih.gov/articles/PMC8276061/. Accessed 2026-08-16.

    Source note · peer-reviewed literature

  9. 9. Wersto RP, Chrest FJ, Leary JF, et al. How to prevent yourself from seeing double. Cytometry Part A. 2020. Peer-reviewed review. https://pmc.ncbi.nlm.nih.gov/articles/PMC7669578/. Accessed 2026-08-16.

    Source note · peer-reviewed literature

  10. 10. Mair F, Hartmann FJ, Mrdjen D, et al. The end of gating? An introduction to high dimensional flow cytometry data analysis. European Journal of Immunology. 2016;46:34-43. Peer-reviewed review. https://pmc.ncbi.nlm.nih.gov/articles/PMC7868168/. Accessed 2026-08-16.

    Source note · peer-reviewed literature

  11. 11. BD Biosciences. Compensation tools on the BD FACSLyric Flow Cytometer. Manufacturer labeling and application guidance. https://www.bdbiosciences.com/content/dam/bdb/marketing-documents/products-pdf-folder/instruments/research-cell-analyzers/Compensation-tools-on-the-BD%20FACSLyric%E2%84%A2-Flow-Cytometer.pdf. Accessed 2026-08-16.

    Source note · manufacturer labeling