Flow cytometry blood cells

Packman, C. H. and Lichtman, M. A. (1990) Activation of neutrophils measurement of actin conformational changes by flow cytometry. Blood Cells 16,193-207. 

Brummendorf TH, Holyoake TL, Rufer N et al. Prognostic implications of differences in telomere length between normal and malignant cells from patients with chronic myeloid leukemia measured by flow cytometry. Blood 2000 95 1883-1890. 

Bender, J.G., Unverzagt, K.L., Walker, D.E., Lee, W., Van Epps, D.E., Smith, D.H., Stewart, C.C., and To, L.B. 1991. Identification and comparison of CD34-positive cells and their subpopulations from normal peripheral blood and bone marrow using multicolor flow cytometry. Blood 77 2591-2596. 

Blood Lymphocytes in Healthy Aged Humans by Flow Cytometry. Immunol Cell Biol 71... 

Several additional instrumental techniques have also been developed for bacterial characterization. Capillary electrophoresis of bacteria, which requires little sample preparation,42 is possible because most bacteria act as colloidal particles in suspension and can be separated by their electrical charge. Capillary electrophoresis provides information that may be useful for identification. Flow cytometry also can be used to identify and separate individual cells in a mixture.11,42 Infrared spectroscopy has been used to characterize bacteria caught on transparent filters.113 Fourier-transform infrared (FTIR) spectroscopy, with linear discriminant analysis and artificial neural networks, has been adapted for identifying foodbome bacteria25,113 and pathogenic bacteria in the blood.5... 

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. 

Flow cytometry is now commonly used in immunotoxicity studies to assess changes in relative frequency and number of lymphoid and myeloid cells in the spleen, lymph nodes, bone marrow and/or peripheral blood of rodents, and in the peripheral blood of humans. A list of selected cell surface markers useful in immunotoxicity studies is shown in Table 7.3. Notably, the majority of available reagents are specific for murine antigens with human reagent availability a close second. Reagents for rat, primate, and... 

Although the concept of flow cytometry originated in studies of blood cells and tumor cells (for review, see Darzynkiewicz et ak, 2004), flow cytometric procedures are now used routinely in disciplines as diverse as immunology, the neurosciences, nutritional sciences, pharmacology, parasitology, and marine biology. This chapter offers an introduction to the theory and practice of analytical flow cytometry, with emphasis on applications in the neurosciences. 

Gati WP, Paterson ARP, Larratt LM, Turner AR, Belch AR 1997. Sensitivity of acute leukemia cells to cytarabine is a correlate of cellular es nucleoside transporter site content measured by flow cytometry with SAENTA-fluorescein. Blood 90 346-53. 

Lombardi, V.R.M., Amado, L., Femandez-Novoa, L., Ftcheverrfa, 1., Seoane, S., Cacabelos, R. (2003) Flow cytometry analysis of CD28-/CD8-I- suppresor cell precursor and CD45RO-I-/ CD4-I- memory T lymphocytes in the peripheral blood of Alzheimer s disease patients. In New trends in Alzheimer- and Parkinson-related disorders, Hanin, 1., Fisher, A., Cacabelos, R. (eds.), Monduzzi Fditore, Bologna, pp. 57-61. 

In another experimental setting in vitro data on the cell surface antigen expression were obtained for PBMCs isolated from chronic lymphocytic leukemia (CLL) patient blood samples and cultured in the presence of DIMS (according to standard procedures). Cultured cells were collected and stained with the corresponding antibodies for detection of the expression of cell surface antigens. Subsequendy they were analyzed by flow cytometry. 

Management depends upon the underlying cause. The cardinal measurement, apart from the preliminary finding of a raised haemoglobin or haematocrit in the blood, is a separate determination of red cell mass and plasma volume using either flow cytometry or the traditional radonuclide methodology. 

Several methods for separating cells have been devised. These include electrophoresisd or use of magnetic microspheres. b Micromanipulation can sometimes be used to select single cells for analysis. The most impressive technique is flow cytometry,ef which is used daily on human blood samples in clinical laboratories. A suspension of cells is passed at a high rate of flow through a narrow capillary of 0.2 mM diameter. The sample stream, which is surrounded by a larger "sheath" stream, has a... 

The second aspect of clinical practice that has led to a reassessment of the nature of flow cytometry is the occasional clinical requirement for rare-event analysis. Methods have been developed, particularly with the use of multiparameter gating, to lower background noise in order to provide increased sensitivity for detection of rare cells. In the clinic, this increased sensitivity translates, for example, into earlier diagnosis of relapse in leukemia, more sensitive detection of fetal-maternal hemorrhage, and better ability to screen leukocyte-reduced blood transfusion products for residual white blood cells. Outside the clinic, these methods for rare-event detection have begun to stretch the limits of research applications as well. 

Fig. 10.6. From Davis BH et al (1998). Detection of fetal red cells in fetomaternal hemorrhage using a fetal hemoglobin monoclonal antibody by flow cytometry. Reprinted with permission from Transfusion 38 749-756, published by the American Association of Blood Banks.

Lymphocyte A lymphocyte is a particular type of white blood cell that is involved in many of an organism s immune responses. Subpopulations of lymphocytes with microscopically identical anatomy can be distinguished because their surface membranes contain different arrays of proteins. The staining of these proteins with fluorescently tagged monoclonal antibodies allows the subpopulations to be enumerated by flow cytometry. 

Sometimes, cellular analysis was performed with cells in a flow stream, which is also termed flow cytometry or FACS in the field of cell biology. For instance, human blood cell (WBC, RBC) rheology was studied in channels fabricated on the Si-Pyrex substrates. The channels were either uncoated or coated with albumin [825]. 

Studies of stem cell progression towards the completely differentiated mature cells have already identified several intermediary precursors, organized in a cascade (Shizuru et al., 2005). The best known and studied cellular differentiation cascade is the hematopoietic system (Figure 20.3). Within hematopoiesis, it is possible to identify many intermediary precursors between the hematopoietic stem cell and mature blood cells. This identification is based mainly on the phenotypic profile of cellular surface proteins, using flow cytometry as the main tool. This is a relatively simple technique that involves coupling a monoclonal antibody (mAb) with a fluorescent marker (fluorochrome). In this way, diverse cellular markers can be combined and thus a cellular subpopulation can be defined, as shown in Figure 20.3. 

Fig. 12.17 Inhibition of phosphorylation of p38 MAP kinase in neutrophils (a, flow cytometry) and cytokine production (c-e, flow-cytometric bead array) in whole human blood stimulated with 100 ng/mL LPS, with either DS-96 or PMB (b) Forward scatter/side scatter profile and the gating for p38 MAPK-negative and positive gates obtained on unstimulated (negative control) and LPS-stimulated (positive control) cells, respectively...

NOVEL PHARMACODYNAMIC MEASURES USING FLOW CYTOMETRY AND BLOOD MONONUCLEAR CELLS AS A SURROGATE FOR TISSUE RESPONSES... 

Chow S, Hedley D, Grom P, Magari R, Jacobberger JW, Shankey TV. Whole blood fixation and permeabilization protocol with red blood cell lysis for flow cytometry of intracellular phosphorylated epitopes in leukocyte subpopulations. Cytometry A 2005 67(1) 4-17. 

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