Flow cytometry cell sorting

Fig. 4. Combined use of flow cytometry/cell sorting and confocal laser scanning microscopy. TNF-a/ActD treated Hepa-1 cells were stained with CMX Rosamine and then analyzed for mitochondrial membrane potential and NAD(P)H fluorescence (A). Cells in different regions of the cytogram were then sorted, and subsequently stained with the DNA fluorochrome Hoechst 33342. These cells were then examined by CLSM. (B), (C), and (D) show three-dimensional reconstructions of nuclei from cells sorted from healthy, early apoptotic, and late apoptotic populations. Whereas healthy cells show normal round nuclei, early and late apoptotic cells show progressive chromatin condensation/margination and nuclear fragmentation.

Flow cytometry (cell sorting, 1972) W first high-throughput screening -> with antibodies follow cellular differentiation separate specific cell types... 

Fig. 8.4. Reprinted with permission of John Wiley Sons, Inc. 1990 from Gray JW, et al. (1990). Quantitative cell-cycle analysis. Melamed MR, et al. (eds). Flow Cytometry and Sorting. New York Wiley-Liss, pp 445-467. The work was performed at the University of California Lawrence Livermore National Laboratory under the auspices of the U.S. Department of Energy.

Stain your cells for flow cytometry or sorting. 

Jefferis, R., Glycosylation of recombinant antibody therapeutics. BiotechnoL Prog., 21, 11, 2005. Marder, et al.. Selective cloning of hybridoma cells for enhanced immunoglobulin production using flow cytometric cell sorting and automated laser nephelometry. Cytometry, 11, 498, 1990. 

Laakso et al. (2001) developed a method for boron determination that uses inductively coupled plasma atomic emission spectrometry and protein removal with trichloroacetic acid before analysis. This method is feasible, accurate, and one of the fastest for boron determination during BNCT and enables a more reliable estimation of the irradiation dose. Yoshida et al. (2002) used flow cytometry to sort the cells by phases, and the boron concentration in each fraction was measured with inductively coupled plasma atomic emission spectroscopy. Obtained results revealed that sodium borocaptate and boronophenylalanine were associated with different rates of boron uptake in different phases. 

Propidium iodide/annexin staining, flow cytometry, and flow-activated cell sorting (FACS) analysis (necrosis, apoptosis, cell cycle status)... 

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). 

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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