Course Outline
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- Flow Cytometry Basics
- 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
- 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
- Conclusion
- 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.