Flow cytometry development

EIA was originally developed as a histological technique to localize specific ceUular sites using the specificity of an immunological reaction (23). The resulting fluorescent antibodies can be detected in animal tissues at levels as low as 1 /tg/mL of body fluid. Eluorophore-labeled antibodies have also been used widely for flow cytometry appHcations using fluorescein antibodies to cell surface markers to detect and quantify specific cells (24). 

We will first summarize the fluorescence and spectroscopic assays that have been developed for the fluorometer and then describe their applications using flow cytometry. We will summarize research which exemplifies the utility of simultaneous measurement of responses and shows how these methods have provided Information about the signal transduction pathways and activation in neutrophils. 

In addition to these time-honoured methods, newer techniques involving bio-luminescense, fluorescent dyes (epifluorescence) and physical methods such as impedance, calorimetry and flow cytometry have been developed. A feature being sought in these methods is rapidity see section 5.6. 

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... 

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... 

Prigione, V., Lingua, G. and Marchisio, V. F. (2004). Development and use of flow cytometry for detection of airborne fungi. Applied and Environmental Microbiology 70 1360-1365. 

One of the first applications developed for flow cytometry was cell cycle analysis.2 There are numerous intercalating fluorescent DNA and RNA staining reagents that can be used to determine the amount of DNA in cells, an indicator of cell cycle stage and progression, as demonstrated in Figure 7.3. Nucleic acid dyes may be selective for DNA... 

In summary, flow cytometry is clearly useful in evaluating macrophages and their role in toxicity. A major advantage of this technology is the rapid and accurate identification of subpopulations of responding cells from within a mixed population. There is no doubt that the utility of flow cytometry will increase in the future as new fluorescence probes are developed that allow investigators to more clearly assess various macrophage characteristics and the response of these cells to xenobiotics. 

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. 

Manetz, T.S. and Meade, B J., Development of a flow cytometry assay for the identification and differentiation of chemicals with the potential to elicit irritation, IgE-mediated, or T cell-mediated hypersensitivity responses, Toxicol. Sci.. 48, 206, 1999. 

Use of Flow Cytometry for Drug Development of Cell Cycle Inhibitor The Example of Edotecarin,... 

Antiproliferative assay, flow cytometry studies, in vivo activities and biomarkers determination represent key tools for profiling the phenotypic response to molecular or pharmacologic perturbation at cellular or tissue levels the study enables therapeutic response irrespective of target activity of topi and taken together, these results suggest that there is a strong potential for edotecarin in clinical [36, 37] development for treatment of human tumors. 

---