Flow cytometry-sorting

Selection is based upon the functional properties of the evolved protein (phenotype). Binding to antigen immobilized in the wells of a microtiter plate or on the surfaces of magnetic beads are frequently used approaches. Flow cytometry sorting systems, some of which can sort up to 108 cells/h, represent a promising new class of selection technique. 

Testing can be performed on genomic DNA isolated from the entire ceU population of the specimen. Alternatively, the sample can be sorted by flow cytometry or immunorosetting to assess engraftment of specific ceil lineages. For example, engraftment of the T-cell versus non-T-cell components can be assessed after flow cytometry sorting based on expression of the CD3 surface molecule, a T-ceU marker. 

J. Porter, C. Edwards, J. A. W. Morgan, and R. W. Pickup, Rapid, automated separation of specific bacteria from lake water and sewage by flow cytometry and cell sorting, Appl. Environ. Microbiol. 59 3327 (1993). 

Darvey, H. M. Kell, D. B. Flow cytometry and cell sorting of heterogenous microbial populations the importance of single cell analysis. Microbiol. Rev. 1997, 60, 641-696. 

Ibrahim, S.F. and van den Engh, G. (2007) Flow cytometry and cell sorting. Advances in Biochemical Engineering/Biotechnology, 106, 19-39. 

The chemical composition of particles can be just as varied as their shape. Commercial particles can consist of polymers or copolymers, inorganic constructs, metals and semiconductors, superparamagnetic composites, biodegradable constructs, and synthetic dendrimers and dendrons. Often, both the composition of a particle and its shape govern its suitability for a particular purpose. For instance, composite particles containing superparamagnetic iron oxide typically are used for small-scale affinity separations, especially for cell separations followed by flow cytometry analysis or fluorescence-activated cell sorting (FACS). Core-shell semiconductor particles, by... 

Radbruch, A., Flow Cytometry and Cell Sorting Springer, Heidelberg (1992). 

A fluorescence-activated cell sorter (FACS) is a flow cytometry instrument used to separate and identify cells in a heterogeneous population. Cell mixtures to be sorted are first bound to fluorescent dyes such as fluorescein or phycoerythrin. The labeled cells are then pumped through the instrument and are excited by a laser beam. Cells that fluoresce are detected, and an electrostatic charge is applied. The charged cells are separated using voltage deflection. 

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). 

Melaned, M. R., Mullaney, P. F and Mendelsohn, M. L. (1979) Flow Cytometry and Sorting. John Willey and Sons, New York. 

Flow cytometry (FCM) is a high-precision technique for rapid analysis and sorting of cells and particles. In theory, it can be used to measure any cell component, provided that a fluorescent tracer is available that reacts specifically and stoichiometrically with that constituent. The technique provides statistical accuracy, reproducibility, and sensitivity. 

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