Multiparameter flow cytometry

Burchiel, S.W. et al., Assessment of immunotoxicity by multiparameter flow cytometry, Fundam. Appl. Toxicol., 38, 38, 1997. 

Applications of Multiparameter Flow Cytometry to the Evaluation of Immunopharmacologic and Immunotoxicologic Actions and Mechanisms... 

Scientific progress has resulted in the continual development and evolution of useful and powerful techniques, such as multiparameter flow cytometry and image analysis, to answer questions of cellular maintenance and function (and the impact chemicals may exert) for the benefit of human health. Multiparameter flow cytometry provides researchers with a capacity to simultaneously examine multiple cellular characteristics on thousands of cells in a relatively short period of time. 

Finally, multiparameter flow cytometry has the potential to improve existing methodologies for the evaluation of immunotoxicological endpoints such as the natural killer cell (NK) assay and the local lymph node assay (LLNA) to evaluate innate immunity and chemical-induced allergic disease. Accepted methodologies typically utilize radio-... 

Analytical flow cytometry offers a rapid and facile means of monitoring cellular receptor content. For example, multiparameter flow cytometry techniques were used to monitor expression of GABAa receptor subunits during neurogenesis in embryonic rat brain (Marie et al., 2001). The content of the cell surface p75 neurotrophin receptor was measured in a heterogeneous population of mouse dorsal root sensory neurons, from which high and low p75 subsets were subsequently isolated by cell sorting (Barrett et al., 1998). 

The following basic protocols may be used alone or combined with other staining procedures in multiparameter flow cytometry experiments. Although they are illustrated with data from cells that proliferate in suspension, these protocols may be easily modified for the analysis of cells isolated from tissues or adherent cells in culture, by incorporating an initial step for the preparation of single cell suspensions. The assays are conducted at room temperature, unless otherwise noted. 

Haskins, J.R., Rowse, P., Rahbari, R. and de La Iglesia, F.A. (2001) Thiazolidinedione toxicity to isolated hepatocytes revealed by coherent multiprobe fluorescence microscopy and correlated with multiparameter flow cytometry of peripheral leukocytes. Archives of Toxicology, 75, 425-438. 

Dive, C., Gregory, C D., Phipps, D J, Evans, D L, Milner, A E, and Wylhe, A H (1992) Analysis and discrimination of necrosis and apoptosis by multiparameter flow cytometry Biochim Biophys Acta 1133, 275—285. 

Fig. 8.14. Reprinted with permission of John Wiley Sons, Inc. 1990 from Darzynkiewicz Z and Traganos F (1990). Multiparameter flow cytometry studies of the cell cycle. Melamed MR, et al. (eds). Flow Cytometry and Sorting. New York Wiley-Liss, pp 469-501.

Leith, C. P., Chen, I-M, Kopecky, K. J., Appelbaum, F. R Head, D. R Godwin, J. E., Weick, J. K., and Willman, C. L. (1995) Correlation of multidrug resistance (MDR1) protein expression with functional dye/drug efflux in acute myeloid leukemia by multiparameter flow cytometry identification of discordant MDR-/ efflux+ and MDR+/efflux cases. Blood 86, 2329-2342. 

Poot, M. and Pierce, R. H. (1999) Detection of changes in mitochondrial function during apoptosis by simultaneous staining with multiple fluorescent dyes and correlated multiparameter flow cytometry. Cytometry 35(4) 311-317. 

Morsi, H. M., Leers, M. P., Radespiel-Troger, M., Bjorklund, V., Kabarity, H. E., Nap, M., and Jager, W. (2000) Apoptosis, bcl-2 expression, and proliferation in benign and malignant endometrial epithelium An approach using multiparameter flow cytometry. Gynecol Oncol. 77(1), 11-17. 

Hewitt C J, Nebe-von Caron G (2001). An industrial application of multiparameter flow cytometry Assessment of cell physiological state and its application to the study of microbial fermentations. C ytometr y 44 179-187. 

Hewitt C J, Nebe-von Caron G, Axelsson B, McFarlane C M, Nienow A W (2000). Studies related to the Scale-up of high-cell-density E. coli fed-batch fermentations using multiparameter flow cytometry Effect of a changing microenvironment with respect to glucose and dissolved oxygen concentration. Biotechnol. Bioengin. 70 381-390. 

For further interpretation of these simulation results, we refer to the experimental observations of Hewitt et a/. [75,76]. These authors have been concerned about studying the influence of the scale of cultivation and, as such, the impact of different intensities of mixing upon the viability off. coli populations during fed-batch fermentations with a constant feeding rate (decreasing glucose concentrations with respect to process time). For this purpose, multiparameter flow cytometry has been applied. With the aid of specific fluorescent dyes, valuable quantitative information on cell physiology and particularly viability could be obtained. The... 

A. Amanullah, C.I. Hewitt, A.W. Nienow, C. Lee, M. Chartrain, B.G. Buckland, S.W. Drew, J.M. Woodley, Measurement of strain-dependent toxicity in the indene bioconversion using multiparameter flow cytometry, Biotechnol. Bioeng. 81 (4) (2003) 405-420. 

A metabolic engineering approach [175] and directed evolution techniques [176] were evaluated to avoid side reactions, block degradative pathways, and enhance the desired reaction (conversion of indene to cxs-amino indanol 137 or ds-indanediol). Multiparameter flow cytometry was used to assess indene toxicity, and it was shown that concentrations up to 025 g/1 of indene (0.037g indene per gram dry cell wt.) in batch bioconversions did not influence reaction rate. Using this information, a single-phase... 

Shapiro, H. M. Multiparameter flow cytometry of bacteria Implications for diagnostics and therapeutics. Cytometry 2001,43,1T3-H6. 

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