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Chapter 11.13 : Whole-Blood Lymphocyte Immunophenotyping Using Cell Surface Markers by Flow Cytometry

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Citation: Leber A. 2016. Whole-Blood Lymphocyte Immunophenotyping Using Cell Surface Markers by Flow Cytometry, p 11.13.1.1-11.13.2.5. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch11.13
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Figures

Image of Figure 11.13.1–1
Figure 11.13.1–1

Light scatter identification of major leukocyte populations. Light scatter properties can be used to identify major leukocyte populations, including neutrophils, monocytes, and lymphocytes. Forward light scatter measurement reflects cell size, and side scatter represents internal complexity.

Citation: Leber A. 2016. Whole-Blood Lymphocyte Immunophenotyping Using Cell Surface Markers by Flow Cytometry, p 11.13.1.1-11.13.2.5. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch11.13
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Image of Figure 11.13.1–2
Figure 11.13.1–2

Lymphocyte immunophenotyping gating strategy. Lymphocytes are selected from a CD45-versus-side scatter two-dimensional plot, and then CD3 is used to discriminate T cells. Non-T cells (CD3) are gated and viewed in a CD19-versus-CD56/16 plot to identify B cells and NK cells, respectively. CD3 T cells are further characterized using a CD4-versus-CD8 plot.

Citation: Leber A. 2016. Whole-Blood Lymphocyte Immunophenotyping Using Cell Surface Markers by Flow Cytometry, p 11.13.1.1-11.13.2.5. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch11.13
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Download as Powerpoint

References

/content/book/10.1128/9781555818814.chap11.13
1. Mandy FF, Nicholson JKA, McDougal JS, Centers for Disease Control and Prevention. 2003. Guidelines for performing single platform absolute CD4+ T-cell determinations with CD45 gating for persons infected with human immunodeficiency virus. MMWR Morb Mortal Wkly Rep 52(RR02):113.
1. Gratama JW, Kraan J, Keeny M, Mandy F, Sutherland DR, Wood BL. 2007. Enumeration on Immunologically Defined Cell Populations by Flow Cytometry. Approved guideline, 2nd ed. CLSI document H42-A2, vol 27, no 16. CLSI, Wayne, PA.
2. Diagnostic Immunology Resource Committee. 2013. FL-A Flow Cytometry Participant Summary. College of American Pathologists Surveys and Anatomic Pathology Education Programs, Northfield, IL.
3. Biosciences BD. 2011. BD Multitest™ 6-color TBNK reagent with Trucount tubes. Product number 23-10834-02. BD Biosciences, San Jose, CA.
4. Biosciences BD. 2010. BD Multitest™ IMK kit. Product number 23-3602-05. BD Biosciences, San Jose, CA.
5. Beckman Coulter. 2012. CYTO-STAT® tetraCHROME CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5 and CYTO-STAT® tetraCHROME CD45-FITC/CD56-RD1/CD19-ECD/CD3-PC5. Product number 4238068-KE. Beckman Coulter, Brea, CA.
6. Beckman Coulter. 2010. Flow-Count fluorospheres. Product number 7507992-MA. Beckman Coulter, Brea, CA.
7. Horowitz GL, Altaie S, Boyd JC, Cerioti F, Garg U, Horn P, Pesce A, Sine HE, Zakowski J. 2008. Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory. Approved guideline, 3rd ed. CLSI document C28-A3, vol 28, no 30. CLSI, Wayne, PA.
8. Denny TN, Louzao R, Wong J, Walker B. 2011. CD4 T cell assessments in evaluation of HIV therapeutics, p 189–204. In Litwin V, Marder P (ed), Flow Cytometry in Drug Discovery and Development. John Wiley & Sons, Inc, Hoboken, NJ.

Tables

Generic image for table
Table 11.13.1–1

Reference ranges

Citation: Leber A. 2016. Whole-Blood Lymphocyte Immunophenotyping Using Cell Surface Markers by Flow Cytometry, p 11.13.1.1-11.13.2.5. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch11.13
Generic image for table
Table 11.13.2–1

Reported results

Citation: Leber A. 2016. Whole-Blood Lymphocyte Immunophenotyping Using Cell Surface Markers by Flow Cytometry, p 11.13.1.1-11.13.2.5. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch11.13

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