Introduction to Flow Cytometry: Blood Cell Identification

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Author: Dana L. Van Laeys, MEd, MLS(ASCP)MBCM, CLSp(MB)
Reviewer: Dr. Linda Miller, PhD I, MBCM(ASCP)SI

No other laboratory method provides as rapid and detailed analysis of cellular populations as flow cytometry, making it a valuable tool for diagnosis and management of several hematologic and immunologic diseases. Understanding this relevant methodology is important for any medical laboratory scientist.
Whether you have no previous experience with flow cytometry or just need a refresher, this course will help you to understand the basic principles, with the help of video tutorials and interactive case studies.

Basic principles include:
  • Immunophenotypic features of various types of hematologic cells
  • Labeling cellular elements with fluorochromes
  • Blood cell identification, specifically B and T lymphocyte identification and analysis
  • Cell sorting to isolate select cell population for further analysis
  • Analyzing and interpreting result reports and printouts

Continuing Education Credits


  • List the appropriate specimen types used for flow cytometric analysis.
  • Describe the steps involved in analyzing samples by flow cytometry.
  • Calculate absolute cell counts from percentage values for flow cytometry analysis.
  • Identify blood cell types associated with respective specific surface markers.
  • Interpret basic flow cytometer instrument analysis printouts.
  • Progress through case studies related to lymphocyte evaluation using flow cytometry while performing final interpretations.

Course Outline

  • Flow Cytometry Basics
      • Flow Cytometry Course Introduction
      • Definitions
  • Flow Cytometry Principles
      • Acceptable Samples
      • Sample Preparation
      • Sample Analysis
      • Sample Analysis Animation
      • Detection of Cellular Characteristics
      • Detection of Intrinsic Cellular Characteristics: Cell Size and Granularity
      • Detection of Extrinsic Cellular Characteristics: Surface Antigens
      • Cell size is indicated by forward scatter and granularity is indicated by side scatter in flow cytometry.
      • Staining Illustration
      • Staining: Monoclonal Antibody Binding
      • T-Helper Cell Example
      • T-Cytotoxic Cell Example
      • Flow Cytometry and Human Immunodeficiency Virus Clinical Application
      • Calculating Absolute Cell Counts
      • Calculating Absolute Cell Counts- continued
      • Which of the following types of lymphocytes express CD4?
  • Identifying and Using Cell Surface Markers
      • Identifying General Cell Types Associated With Surface Markers
  • Interpreting Instrument Printouts
      • Cytogram for Cellular Scatter: Intrinsic Characteristics
      • White Blood Cells and Cytogram for Cellular Scatter
      • Extrinsic Cell Characteristics
      • Which of these white blood cell populations would have the MOST side scatter when analyzed using flow cytometry?
      • Gating: Selecting the Population of Interest
      • Using Cell Surface Markers as Tools For Diagnosis
      • Diagnostic Process
  • Review of Flow Cytometry Principles
      • Principles of Flow Cytometry
  • Case Study #1
      • Case One
      • Scatter and CD45
      • What cell population is gated in this peripheral blood sample case analyzed by flow cytometry?
      • CD19 and CD20
      • It has already been established that the gated cell population is lymphocytic in nature. CD19 and CD20 cell surface antigens both appear on what type ...
      • CD10, HLA-DR, Kappa, and Lambda
      • If 100% of the cells in the gated lymphocyte population are positive for CD45 (i.e., they are leukocytes) and 7% are B cells (represented by CD19 and ...
      • T Cell Markers
      • All helper cells are T cells. Consequently, in a normal lymphocyte population, it is reasonable to expect all CD4-positive cells to be CD3-positive as...
      • Final Interpretation of Case #1
  • Case Study #2
      • Case Two
      • Scatter and Gating
      • T Cell Analysis
      • B Cell Analysis and Final Interpretation of Case Two
  • Conclusion
      • Conclusion
  • References
      • References

Additional Information

Level of instruction: Intermediate

Intended Audience: Medical laboratory scientists, medical technologists, and technicians. This course is also appropriate for medical laboratory science students and pathology residents.
Author information: Dana L. Van Laeys, MEd, MLS(ASCP)MBCM is the Education Coordinator for Molecular Diagnostics and Immunology  in the Clinical Laboratory Science Program at Saint Luke’s Hospital in Kansas City, Missouri. She has 14 years of experience in molecular diagnostics and flow cytometry. She has been a presenter at ASM, CLEC, and AABB conferences. Ms. Van Laeys holds a Masters in Adult Education and Distance Learning.
Reviewer information: Dr. Linda Miller, PhD I, MBCM(ASCP)SI, is an Assistant Editor for the Immunohematology section of Lab Medicine. She received her BS degree in Biology from Syracuse University and her PhD in Immunology from SUNY Upstate Medical University. She holds certifications as a Technologist in Immunology, Specialist in Immunology, and Technologist in Molecular Biology from the ASCP. Dr. Miller is a professor of Clinical Laboratory Science at Upstate Medical University and the Director of the Medical Biotechnology program in the same department. There, she teaches courses in Immunology, Human Genetics, Molecular Methods, and Medical Biotechnology. She has served as a member of the ASCP Immunology Exam Committee and the ASCP Lab Q Editorial Board as Associate Editor of Clinical Immunology. Dr. Miller has written numerous continuing education exercises and book chapters in clinical immunology and is co-author of the 4th edition of the textbook, Clinical Immunology and Serology: A Laboratory Perspective (FA Davis).
Course description: This course describes the most basic principles of flow cytometry, providing an overview and introduction to this useful tool. The focus of the course is blood cell identification, specifically B and T lymphocyte identification and analysis.

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