Flow cytometry, applications

FIA was originally developed as a histological technique to localize specific cellular 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. Fluorophore-labeled antibodies have also been used widely for flow cytometry applications using fluorescein antibodies to cell surface markers to detect and quantify specific cells (24). 

Sundararajan N, Pio M, Lee L, Berlin A (2004) Three-dimensional hydrodynamic focusing in polydimethylsiloxane (PDMS) microchannels. J Microelectromech Syst 13(4) 559-567 Yang R, Feeback D, Wang W (2005) Microfabrication and test of a three-dimensional polymer hydro-focusing unit for flow cytometry applications. Sens Actuators A Phys 118(2) 259-267... 

The membrane potential of individual cells can be monitored with a fluorescence microscope. For this purpose, however, it is preferable to use a permeable redistribution dye with spectral characteristics that have minimal environmental sensitivity. Thus, the fluorescence intensity will reflect the degree of Nemstian accumulation of dye only and can, therefore, be readily interpreted. The plasma membrane potential can be distinguished from the organelle membrane by simply using the microscope to identify appropriate regions of the cell (44). Rhodamine-123 (Chart III) was introduced as a mitochondrial stain by Chen and co-workers (45-47) it has been used largely in qualitative studies of mitochondrial membrane potential and has been especially effective in flow cytometry applications. 

Much of the early work in this area was done for flow cytometry applications using embedded or proximity fiber systems. These devices typically measured the counts and velocities of microspheres in liquids. These systems are able to detect 10 pm spheres at a rate of 65 particles per second (Table 1, 2nd row) [12]. 

Electrostatic manipulation of droplets has been utilized for ink-jet printing, ESI mass-spectrometry and flow cytometry applications. In contrast to these droplet-in-air applications, electrowetting involves the electrostatic manipu-... 

Silica-fluorescent dye beads have not been utilized as much as the PS-based ones. As pointed out recently in our efforts these are easily prepared in abundance, and have optical and chemical features that lend them suitable for flow cytometry applications. The limiting feature—concentration quenching—of any fluorophore-bead system is illustrated in Fig. 11 for rhodamine 6G in silica. Here, successively higher concentrations of R6G in a layer of silica-R6G around the same core silica showed quenching of R6G emission from the increase to a maximum at S and then decrease in intensity for samples S-1 to S-6 shown in Fig. 11. 

Ronot X, Paillasson S and Muirhead, KA (1996) Assessment of cell viability in mammalian cell lines. In Al-Rubeai M and Emery AN (eds) Flow Cytometry Applications in Cell Culture, pp 177-192. New York Marcel Dekker. 

Telford, W. G. Cox, W. G. Singer, V. L. Detection of endogenous and antibody-conjugated alkaline phosphatase with ELF-97 phosphate in multicolor flow cytometry applications. Cytometry 2001, 43, 117-125. 

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. 

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... 

The CTAD additive mixture has found application in the monitoring of heparin therapy by either the chromogenic substrate assay or the APTT and in the measurement of platelet markers such as P-selectin (CD62) by flow cytometry (108, 109). 

Gunasekera,T. S. Veal, D. A. Attfleld,P. V. Potential for broad applications of flow cytometry and fluorescence techniques in microbiological and somatic cell analyses of milk. Int. J. Food Microbiol. 2003, 85, 269-279. 

Most recent scientific applications involve the determination of direct relationships between input parameters and a known target response. For example, Santana and co-workers have used ANNs to relate the structure of a hydrocarbon to its cetane number,4 while Berdnik s group used a theoretical model of light scattering to train a network that was then tested on flow cytometry data.5... 

While more commonly used to count or otherwise characterize cells for medical applications, Coulter Counters and flow cytometry technique can also be applied to the analysis of pollen grains in allelopathic studies. They are quite useful in determining the size and number of pollen grains. The technique is often used for assessing the production and size of pollen from the originating individual rather than how much was transferred to heterospecific stigma, as would be needed in a basic assessment of potential allelopathic interactions. 

Immunotoxicity Testing Committee, Application of flow cytometry to immunotoxicity testing summary of a workshop report, ILSI/HESI, Washington D.C., 1999. 

Burchiel, S.W., et al., Uses and future applications of flow cytometry in immunotoxicity testing, Methods, 19, 28, 1999. 

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

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... 

A series of silica nanoparticles doped with cyanine dye DY-635 (Dyomics) were also prepared, characterized, and investigated in flow cytometry and fluorescence imaging applications [77]. Also these dye-nanoparticles demonstrate high, stable, and tunable fluorescence intensity and are useful for multicolor detection. 

Likewise, the determination of DNA index by flow cytometry and the ability to assess the fraction of cells in the S phase also have applications in oncology, as was discussed earlier. 

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