DNA analysis, by flow cytometry

Deitch, A. D., Law, H., and DeVere White, R. (1992) A stable propidinm iodide staining procednre for flow cytometry. J. Histochem. Cytochem. 30,967-972. Taylor, I. W. (1980) A rapid single step staining techniqne for DNA analysis by flow microflnorimetry. J. Histochem. Cytochem. 28,1021-1024. 

In addition to the use of electrophoresis for identification of the characteristic DNA ladders, DNA content analysis by flow cytometry is often employed to identify cells undergoing DNA fragmentation. This method is based on the fact that permeabUized apoptotic cells release... 

In addition to the use of electrophoresis for identification of the characteristic DNA ladders, DNA content analysis by flow cytometry is often employed to identify cells undergoing DNA fragmentation. This method is based on the fact that permeabilized apoptotic cells release fragmented DNA, resulting in a DNA content that is less than that in live diploid cells. Sohn et al. [10] developed a novel microfluidic technique - capacitance cytometry -that can be used to quantify the polarization response of... 

Miller BM and Nusse N (1993). Analysis of micronuclei induced by 2-chlorobenzylidene malononitrile (CS) using fluorescence in situ hybridization with telemetric and centrometric DNA probes and flow cytometry. Mutagenesis, 8, 35-41. 

SMOKELESS TOBACCO AND APOPTOSIS IN HUMAN ORAL KERATINOCYTES BY FLOW CYTOMETRY AND DNA CELL CYCLE ANALYSIS... 

To get DNA histograms, after separating by tripsinization, cells were fixed with 20% ethanol. They were treated with 0.25% ribonuclease for 1 hr followed by the staining of intranuclear DNA with 0.005% propidium iodide. DNA histograms were drawn by flow cytometry (Cytofluorograf 50H, Ortho Co.) followed by cell cycle analysis. ... 

M. Estrada, P. Henriksen, J.M. Gasol, E.O. Casamayor and C. Pedros-Alio, Diversity of planktonic photoautotrophic microorganisms along a salinity gradient as depicted by microscopy, flow cytometry, pigment analysis and DNA-based methods. FEMS Microbiol. Ecol. 49 (2004) 281-293. 

Flow cytometry (FCM) is widely used for exploring mechanism of action of compounds that compromise proliferation since it is rapid, accurate and usable for any cellular context [5], In this chapter we want to point out technical and strategic aspects of use of FCM for cell cycle studies of a putative anticancer agent. As an example we used Edotecarin, a topi inhibitor, firstly evaluating proliferation outcome and classical DNA content analysis by propidium iodide, and then since the compound treatment produced cell cycle perturbation difficult to interprete, a two-parametric analysis by 5-bromo-deoxyuridine (BrdU) was applied for separating cell cycle phases. Moreover we put our efforts into identifing specific cell cycle arrest not easily demonstrable by previously described methods, through the use of in vitro kinetics ( pulse and chase ). Finally, in vivo assessment of efficacy and biomarkers modulation after treatment was analyzed. 

The analysis of cell-cycle progression was one of the earliest applications of flow cytometry (for review, see Darzynkiewicz et al., 2004). In this assay, fluorescence signals from cells stained with DNA-binding fluorochromes are plotted as DNA content histograms that may be analyzed by using histogram deconvolution software to quantify cell-cycle phase distributions (Rabinovitch 1994). Fluorochromes that are useful for this purpose are the plasma membrane-impermeant DNA stains, propidium iodide (PI),... 

Videl, L. L. and Christensen, I. J. (1990) A review of techniques and results obtained in one laboratory by an integrated system of methods designed for routine clinical flow cytometric DNA analysis. Cytometry 11, 753-770. 

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