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Biomedical Uses of Flow Cytometry

  • James L. Weaver
  • Maryalice Stetler-Stevenson
Protocol
  • 3.1k Downloads
Part of the Springer Protocols Handbooks book series (SPH)

1. Definition of Flow Cytometry

Flow cytometry is literally measuring cells or particles while they are moving in a liquid. More specifically, a suspension of single cells is labeled with one or several fluorescent labels. In the machine, the cells are constrained into single file. These cells pass through one or more laser beams to excite the fluorescent labels. The light emitted from the fluorescent labels is collected, separated, measured, and the resulting data transmitted to the computer controlling the instrument (Fig. 57.1). In addition, narrow angle and 90° light scatter from the laser beam are measured and the data are also sent to the computer. All of these values are recorded as correlated measurements for each cell separately. The data are displayed in the computer as single parameter histograms or two parameter plots (Fig. 57.2). The software allows populations to be identified and specific subpopulations selected for further analysis. The number and fraction of cells in...

Keywords

Flow Cytometry Acute Myeloid Leukemia Hairy Cell Leukemia Autoimmune Hemolytic Anemia Fetal Hemoglobin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Shapiro H (2003) History. In: Practical flow cytometry, 4th edn, John Wiley, Hoboken, New JerseyCrossRefGoogle Scholar
  2. 2.
    Herzenberg LA, Parks D, Sahaf B, Perez O, Roederer M, Herzenberg LA (2002) The history and future of the fluorescence activated cell sorter and flow cytometry: a view from Stanford. Clin Chem 48:1819–1827PubMedGoogle Scholar
  3. 3.
    Jaffe ES, Harris NL, Stein H, Vardiman JW (eds) (2001) WHO classification, pathology and genetics tumors of haematopoietic and lymphoid tissues. IARC, Lyon, FranceGoogle Scholar
  4. 4.
    Tbakhi A, Edinger M, Myles J, Pohlman B, Tubbs RR (1996) Flow cytomet-ric immunophenotyping of non-Hodgkin's lymphoma and related disorders. Cytometry 25:113–124PubMedCrossRefGoogle Scholar
  5. 5.
    Weir EG, Borowitz MJ (2001) Flow cytometry in the diagnosis of acute leukemia. Semin Hematol 38:124–138PubMedCrossRefGoogle Scholar
  6. 6.
    Fukushima PI, Nguyen PK, O'Grady P, Stetler-Stevenson M (1996) Flow cytomet-ric analysis of kappa and lambda light chain expression. Commun Clin Cytometry, 26:243–252CrossRefGoogle Scholar
  7. 7.
    Li S, Eshleman JR, Borowitz MJ (2002) Lack of surface immunoglobulin light chain expression by flow cytometric immunophenotyping can help diagnose peripheral B cell lymphoma. Am J Clin Pathol 118:229–234PubMedCrossRefGoogle Scholar
  8. 8.
    Kuchnio M, Sausville EA, Jaffe ES, Greiner T, Foss FM, McClanahan J, Fukushima P, Stetler-Stevenson MA (1994) Flow cytometric detection of neoplas-tic T cells in patients with mycosis fungoides based upon levels of T-cell receptor expression. Am J Clin Pathol 102:856–860PubMedGoogle Scholar
  9. 9.
    Edelman J, Meyerson HJ (2000) Diminished CD3 expression is useful for detecting and enumerating Sezary cells. Am J Clin Pathol 114:467–477PubMedGoogle Scholar
  10. 10.
    Gorczyca W, Weisberger J, Liu Z, Tsang P, Hossein M, Wu CD, Dong Wong JYL, Tugulea S, Dee S, Melamed MR, Darzynkiewicz Z (2002) An approach to diagnosis of T-cell lymphoproliferative disorders by flow cytometry. Cytometry (Clin Cytometry) 50:177–190CrossRefGoogle Scholar
  11. 11.
    van den Beemd RB, Boor PPC, van Lochem EG, Hop WCJ, Langerak AW, Wolvers-Tettero ILM, Hooijkaas H, van Dongen JJM (2000) Flow cytometric analysis of the V beta repertoire in healthy controls. Cytometry 40(4):336–345PubMedCrossRefGoogle Scholar
  12. 12.
    Lima M, Almeida J, Santos AH, dos Anjos Teixeira M, Alguero MC, Queiros ML, Balanzategui A, Justica B, Gonzalez M, San Miguel JF, Orfao A (2001) Immunophenotypic analysis of the TCR-V repertoire in 98 persistent expansions of CD3+/TCR large granular lymphocytes. Am J Pathol 159:1861–1868PubMedCrossRefGoogle Scholar
  13. 13.
    Weir EG, Borowitz MJ. (2001) Flow cytometry in the diagnosis of acute leukemia. Semin Hematol 38:124–138PubMedCrossRefGoogle Scholar
  14. 14.
    Borowitz MJ, Guenther KL, Shults KE, Stelzer GT (1993) Immunophenotyping of acute leukemia by flow cytometric analysis: use of CD45 and right angle light scatter to gate on leukemic blasts in three color analysis. Am J Clin Pathol 100:534–540PubMedGoogle Scholar
  15. 15.
    Ranier R, Hodges I, Stelzer G (1995) CD45 gating correates with bone marrow differential. Cytometry 22:139–145CrossRefGoogle Scholar
  16. 16.
    Stetler-Stevenson M, Schrager JA (2006) Flow cytometry In: Keren DF, McCoy JP, CareyJL (eds), Clinical Diagnosis, 4th edn., ASCP, Chicago 129–167Google Scholar
  17. 17.
    Garratty G, Arndt PA (1999) Applications of Flow Cytofluorometry to Red Blood Cell Immunology. Cytometry 38:259–267PubMedCrossRefGoogle Scholar
  18. 18.
    Davis BH, Olsen S, Bigelow NC, Chen JC (1998) Detection of fetal red cells in fetomaternal hemorrhage using a fetal hemoglobin monoclonal antibody by flow cytometry. Transfusion 38:749–756PubMedCrossRefGoogle Scholar
  19. 19.
    Ault KA (2001) The clinical utility of flow cytometry in the study of platelets. Semin Hematol 38:160–168PubMedCrossRefGoogle Scholar
  20. 20.
    Davis B, Bigelow N (1999) Indirect immunoplatelet counting by flow cytometry as a reference method for platelet count calibration. Lab Mematol 5:15–21Google Scholar
  21. 21.
    Tschoepe D, Schultheiss HP, Kolarov P, Schwippert B, Dannehl K, Nieuwenhuis HK, Kehrel B, Strauer B, Gries FA (1993) Platelet membrane activation markers are predictive for increased risk of acute ischemic events after PTCA. Circulation 88:37–42PubMedGoogle Scholar
  22. 22.
    His ED (2000) Paroxysmal nocturnal hemaglobinuria testing by flow cytometry – Evaluation of the REDQUANT and CELLQUANT kits. AJCP 114:798–806Google Scholar
  23. 23.
    Hernandez-Campo PM, Martin-Ayuso M, Almeida J, Lopez A, Orfao A (2002) Comparative analysis of different flow cytometry-based immunophenotypic methods for the analysis of CD59 and CD55 expression on major peripheral blood cell subsets. Cytometry 50:191–201PubMedCrossRefGoogle Scholar
  24. 24.
    Horsbugh T, Martin S, Robson AJ (2000) The application of flow cytometry to histocompatibility testing. Transplant Immunol 8:3–15CrossRefGoogle Scholar
  25. 25.
    Kerman RH, Gebel H, Bray R, Garcia C, Sahai R, Gibson H, Radovancevic B, Knight R, Katz S, VanBuren C, Kahan B. (2002) HLA antibody and donor reactivity define patients at risk for rejection or graft loss. Tissue Antigens 59:60–660CrossRefGoogle Scholar
  26. 26.
    Bryan CF, McDonald SB, Baier KA, Luger AM, Aeder MI, Murillo D, Muruve NA, Nelson PW, Shield CF 3rd, Warady BA (2002) Flow cytometry beads rather than the antihuman globulin method should be used to detect HLA Class I IgG antibodies (PRA) in cadaveric renal regraft candidates. Clin Transpl 16:15–23CrossRefGoogle Scholar
  27. 27.
    Davis BH, Olsen SH, Ahmad E, Bigelow NC (2006) Neutrophil CD64 is an improved indicator of infection or sepsis in Emergency department patients. Arch Pathol 130:654–661Google Scholar
  28. 28.
    Mandy F, Nicholson J, Autran B, Janossy G (2002) T-cell subset counting and the fight against AIDS: reflections over a 20-year struggle. Cytometry (Clin Cytometry) 50:39–45CrossRefGoogle Scholar
  29. 29.
    Giorgi JV, Lyles RH, Matud JL, Yamashita TE, Mellors JW, Hultin LE, Jamieson BD, Margolick JB, Rinaldo Jr CR, Phair JP, Detels R. (2002) Predictive value of immunologic and virologic markers after long or short duration of HIV-1 infection. J Acq Imm Def Dis 29:346–355Google Scholar
  30. 30.
    O'Gorman MR, Scholl PR (2002) Role of flow cytometry in the diagnostic evaluation of primary immunodeficiency disease. Clin Appl Immunol Rev 2:321–335.CrossRefGoogle Scholar
  31. 31.
    Bleesing JJH. (2003) Autoimmune lymphoproliferative syndrome (ALPS). Curr Pharm Design 9:265–278CrossRefGoogle Scholar
  32. 32.
    Ebo DG, Sainte-Laudy J, Bridts CH, Mertens CH, Hagendorens MM, Schuerwegh AJ, De Clerk LS, Stevens WJ (2006) Flow-assisted allergy diagnosis: current and future perspectives. Allergy 61:1028–1039PubMedCrossRefGoogle Scholar
  33. 33.
    Bienvenu J, Monneret G, Fabien N, Revillard JP. (2000) The clinical usefulness of the measurement of cytokines. Clin Chem Lab Med 38:267–285PubMedCrossRefGoogle Scholar
  34. 34.
    Tricot G, Jagannath S, Vesole D, Nelson J, Tindle S, Miller L, Cheson B, Crowley J, Bariogi B (1995) Peripheral blood stem cell transplants for multiple myeloma: identification of favorable variables for rapid engraftment in 225 patients. Blood 85:588–596PubMedGoogle Scholar
  35. 35.
    Weaver CH, Hazelton B, Birch R, Palmer P, Allen C, Schwartzberg L, West W (1995) An analysis of engraftement kinetics as a function of the CD34 content of peripheral blood progenitor cell collections in 692 patients after the administration of myeloablative chemotherapy. Blood 86:3961–3969PubMedGoogle Scholar
  36. 36.
    Sutherland DR, Keating A, Nayar R, Anania S, Stewart AK (1994) Sensitive detection and enumeration of CD34+ cells in peripheral blood and cord blood by flow cytometry. Exp Hematol 22:1003–1010PubMedGoogle Scholar
  37. 37.
    Gratama JW, Orfao A, Barnett D, Brando B, Huber A, Janossy G, Johnsen HE, Keeney M, Marti GE, Preijers F, Rothe G, Serke S, Sutherland DR, Van der Schoot CE, Schmitz G, Papa S (1998) Flow cytometric enumeration of CD34(+) hemat-opoietic stem and progenitor cells. Cytometry 34:128–142PubMedCrossRefGoogle Scholar
  38. 38.
    Sutherland DR, Anderson L, Keeney M, Nayar R, Chin-Yee I (1996) The ISHAGE guidelines for CD34 cell determination by flow cytometry. J Hematother 5:213–226PubMedCrossRefGoogle Scholar
  39. 39.
    Barnett D, Granger V, Kraan J, Whitby L, Reilly JT, Papa S, Gratama JW (2000) Reduction of intra and interlaboratory variation in CD34 positive stem cell enumeration by the use of stable test material, standard protocols and targeted training. Br J Haematol 108:784–792PubMedCrossRefGoogle Scholar
  40. 40.
    Goddard G, Martin JC, Naivar M, Goodwin PM, Graves SW, Habbersett R, Nolan JP, Jett JH (2006) Single particle high resolution spectral analysis flow cytometer. Cytometry 69A:842–851CrossRefGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • James L. Weaver
    • 1
  • Maryalice Stetler-Stevenson
    • 2
  1. 1.Department of Applied Pharmacology ResearchCDERWhite Oak
  2. 2.Laboratory of PathologyNCI, NIHBethesda

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