Flow cytometric methods

Derer, M., Walker, C., Kristensen, F., and Reinhardt, M. C. (1983) A simple and rapid flow cytometric method for routine assessment of baker s yeast uptake by human polymorphonuclear leukocytes. J. Immunol. Methods 61, 359-365. 

Figure 17.10 Flow cytometric method does not differentiate between nucleated erythroid cells and reticulocytes, on some instruments. Non-nucleated e throid cells were isolated using cellulose fractionation. Part of the sample was stained with Wright-Giemsa and reticulocytes and mature nucleated cells quantified by trained medical technologists.

This chapter describes a simple immunoperoxidase method for detection of PCNA in tissue sections. A flow cytometric method for demonstrating PCNA in single cells is given in Chapter 36. 

Appropriate application of flow cytometric methods allows cellular fluorescence to be measured and calibrated accurately. In this way, flow cytometric analysis of target antigen density can be undertaken. 

Cole, A. T., Garlick, N M, Galvin, A M., Hawkey, C J., and Robins, R A (1995) A flow cytometric method to measure shape change of human neutrophils. Clin Set 89,549-554... 

Flow Cytometric Methods of Analyzing Apoptotic Cells... 

Currently flow-cytometric methods are also under development to examine effects of various inhibitors of PARP (poly(ADP-ribose)polymerase) that are in preclinical development or clinical trials and use native or stimulated PARP activity in peripheral blood mononuclear cells. 

One such method is fhe microsphere-based xMap technology (Luminex, Austin, Texas, USA), which is a multiplex flow cytometric method based on antibodies coupled to spectrally specific fluorescent microspheres (Vignali, 2000). Another fluorescent reporter antibody binds the protein captured on the microspheres. 

Belloc, F, Dumain, R, Boisseau, M. R., Jalloustre, C., Reiffers, J., Bernard, P. and Lacombe, R (1994). A flow cytometric method using Hoechst 33342 and pro-pidium iodide for simultaneous cell cycle analysis and apoptosis determination in unfixed cells. Cytometry 17, 59-65. 

The flow cytometric method may not be suitable for potential antiviral compounds that significantly alter the host cell membranes because this may alter the appearance (size or granularity) or permeability and hence the staining of the cells. 

Flow cytometric methods can be used to quantify apoptosis among a population of dispersed cells. Two approaches may be used (1) detection of apoptotic cells as indicated by ploidy status (apoptotic cells exhibit hypoploidy due to DNA degradation) (2) detection of apoptotic cells after fluorescent tagging of phosphatidylserine residues in the outer leaflet of the plasma membrane. This approach is limited by the necessity to obtain relatively pure populations of dispersed cells, and as such is not amenable to all tissue types. 

Makrigiorgos, G. M., Kassis, A. I., Mahmood, A., Bump, E. A., and Savvides, P. (1997) Novel fluorescein-based flow-cytometric method for detection of lipid peroxidation. Free Radic. Biol. Med. 22(1-2), 93-100. 

Endresen, P. C., Prytz, P. S., and Aarbakke, J. (1995) A new flow cytometric method for discrimination of apoptotic cells and detection of their cell cycle specificity through staining of F-actin and DNA. Cytometry 20(2), 162-171. 

Poot, M., Verkerk, A., Koster, J. F and Jongkind, J. F. (1986) De novo synthesis of glutathione in human fibroblasts during in vitro ageing and in some metabolic diseases as measured by a flow cytometric method. Biochim. Biophys. Acta 883(3), 580-584. 

Bochner BS, McKelvey AA, Schleimer RP, Hildreth JE, MacGlashan DW Jr (1989) Flow cytometric methods for the analysis of human basophil surface antigens and viability. J Immunol Methods 125 265-271... 

Steensma DP, Timm M, WitzigTE. Flow cytometric methods for detection and quantification of apoptosis. Methods Mol Med 2003 85 323-332. 

Bracher M, Gould HJ, Sutton BJ, Dombrowicz D, Karagiannis SN. Three-colour flow cytometric method to measure antibody-dependent tumor cell killing by cytotoxicity and phagocytosis. Journal of Immunological Methods. 2007 323(2) 160-71. 

Boyd, A. R., Gunasekera, T. S., Attfleld, P. V., Simic, K., Vincent, S. F., Veal, D. A. (2003). A flow-cytometric method for determination of yeast viability and cell number in a brewery. FEMS Yeast Research, 3, 11-16. 

To date, the activities of all research groups that aim to improve methods for the determination of GSH and related compounds are greatly increased. To overcome the difficulties associated with sample pretreatment and conversion of the analytes to detectable derivatives, the application of automated methods, such as flow injection (FI) or sequential injection (SI) techniques, is especially promising and provides viable alternative for the determination of GSH and GSSG in a variety of sample matrixes [60]. The reported FI and SI methods for the determination of GSH are based on UV-Vis [61-64], calorimetric [65], chemiluminescence (CL) [66-69], and electrochemical [70-72] detection. In addition, flow cytometric methods with fluorometric detection that allow rapid measurement of cellular GSH content in clinical samples, namely in tumor tissue, would be useful for biochemical analysis [73,74]. 

Pilas, B. Durack, G. A flow cytometric method for measurement of intracellular chloride concentration in lymphocytes using the halide-specific probe 6-methoxy-AT-(3-sulfopropyl) quinolinium (SPQ). Cytometry 1997, 28, 316-322. 

Bunthof, C. J. Abee, T. Development of a flow cytometric method to analyze subpopulations of bacteria in probiotic products and dairy starters. Appl. Environ. Microbiol. 2002,68, 2934—2942. 

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