Immunophenotyping, flow cytometry

Flow cytometry has been at the forefront of immunophenotyping (see Chapter 3.2). Immunophenotyping is the term used to describe the application of fluorescently tagged antibodies to discriminate and quantitate the various constituent cells of the immune system by flow cytometry. Immunophenotyping cells on the basis of cell surface markers can be thought of as the flrst generation application of flow cytometry in immunotoxicology. 

Kothari R, Cualing H, Balachander T. Neural network analysis of flow cytometry immunophenotype data. IEEE Trans Biomed Eng 1996 43(8) 803-810. 

Immunophenotyping by flow cytometry has taken on an increasingly important role in the diagnosis of leukemia. Owing to the ease of application, sensitivity, and quantifiable results, flow cytometry is the preferred method for lineage assignment.7 This approach takes advantage of the development of monoclonal antibodies... 

Follow-on studies are also recommended as needed. These include determination of potential test article effects on blood or tissue immunophenotypes (by flow cytometry or immunohistochemistry), natural killer cell, macrophage, or neutrophil function, host resistance to infection or tumors, and cell-mediated immunity. The important issue in all of these guidelines is this do not ignore signs of immunotoxicity, and assess these findings when observed. 

Lyons AB, Samuel K, Sanderson A, Maddy AH. 1992. Simultaneous analysis of immunophenotype and apoptosis of murine thymocytes by single laser flow cytometry. Cytometry 13 809-821. 

Comparison of Techniques of Immunophenotypic Analysis Flow Cytometry vs. Immunohistochemistry... 

Baumgarth, N. and Roederer, M. 2000. A practical approach to multicolor flow cytometry for immunophenotyping. J Immunol Methods 243 77-97. 

Immunophenotyping either by immunohistochemistry in lymphoid tissues or flow cytometry in blood samples to enumerate changes in specific leukocyte populations. 

Loken, M. R., Brosnan, J M., Bach, B. A., and Ault, K. A (1990) Establishing optimal lymphocyte gates for immunophenotyping by flow cytometry Cytometry... 

Kim GGG, Donnenberg VSV, Donnenberg ADA, Gooding WW, Whiteside TLT (2007) A novel multiparametric flow cytometry-based cytotoxicity assay simultaneously immunophenotypes effector cells comparisons to a 4 h 51Cr-release assay. J Immunol Methods 325 16-66... 

Bigos M, Baumgarth N, Jager GC, Herman OC, Nozaki T, Stovel RT, Parks DR, Herzenberg LA (1999) Nine color eleven parameter immunophenotyping using three laser flow cytometry. Cytometry 36 36-45... 

A number of methods for the study of apoptosis and necrosis by flow cytometry have been developed (see review by Steensma et al., 2003). Unfortunately, none of the methods are rigorously specific for apoptosis and many show poor overall specificity for cell death or cannot discriminate between the terminal stages of apoptosis and necrosis. Moreover, some of the fluorochromes used to detect apoptosis have emission spectra that fully overlap the spectra of those typically used for immunophenotyping or have a broad emission spectrum that makes compensation in multi- or polychromatic flow cytometry difficult, if not impossible. Thus, the preferred method would be one that measures two aspects of apoptosis, can discriminate between apoptotic and dead cells, and uses fluorochromes that allow simultaneous immunophenotyping. One such method is the annexin-V/7-amino-actinomycin-D assay described by Merchant et al. (2001) that is available in kit from BD Biosystems (San... 

Treatment results in childhood acute leukemia continue to be improved due to new therapeutic and diagnostic methods. An optical microscopy method with specific staining allows to classify acute leukemia into lymphoblastic and myeloid types and discriminate its subtypes according to principals of FAB-classification. A flow cytometry method with monoclonal antibodies staining allows us to discriminate different immunophenotypic subtypes of acute... 

Flow cytometry is commonly used in the clinical diagnostics, for immunophenotyping leukemia, counting stem cells for optimizing autologous transplants for the treatment of leukemia, counting CD4-1-/ CD8-I- lymphocytes in HIV-infected patients to determine the progression of the disease, and the analysis of stained DNA from solid tumors [23],... 

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