Analysis flow cytometry

MCF-7 cells exposed to 50 nM TSA were in G1 phase ( 70%). However, cells treated with 1 J,M TSA arrested cells predominantly in G2/M phase, suggesting a dose-dependent fashion. With either dose, cells accumulated least in the S phase. In addition to accumulation of hyperacetylated nucleosome core histones, TSA enhanced p21 expression. Therefore, flow cytometry (FACS) offers an efficient tool for analyzing the action of HDACIs in cell proliferation. This method is particularly useful for application to clinical samples, where cell numbers may be small. The effects of HDACIs on the cell cycle are dose-dependent. 

Furthermore, since the cell growth arrest is often linked to cell death. The annexin V staining positive cell or the amount of DNA fragmentation assessed by TUNEL and FACS analysis has been interpreted as indicative of apoptosis. The HDACI-induced apoptosis can also be determined by Western blotting of target proteins, detection of mitochondrial membrane potentials, activation of caspases and their substrate cleavages in a dose- and time-dependent manner. 

Inhibition of HDACs has become an interesting approach for anticancer therapy. Therefore, two approaches, gene knockdown in cells and knockout in animals, have been developed and widely used for functional studies of specific HDAC isoforms. 

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Fig. 3. Flow cytometry analysis. CellQuest software. The fluorescence of 50,000 cells is measured.

Anderson MT, Baumgarth N, Haugland RP et al (1998) Pairs of violet light excited fluorochromes for flow cytometry analysis. Cytometry 33 435-444... 

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... 

OXIN O partially inhibited ADP-induced platelet aggregation in vitro. Flow cytometry analysis of PRP showed that OXINO decreased ADP-induced CD62P expression and fibrinogen binding on platelets [45]. 

Lombardi, V.R.M., Amado, L., Femandez-Novoa, L., Ftcheverrfa, 1., Seoane, S., Cacabelos, R. (2003) Flow cytometry analysis of CD28-/CD8-I- suppresor cell precursor and CD45RO-I-/ CD4-I- memory T lymphocytes in the peripheral blood of Alzheimer s disease patients. In New trends in Alzheimer- and Parkinson-related disorders, Hanin, 1., Fisher, A., Cacabelos, R. (eds.), Monduzzi Fditore, Bologna, pp. 57-61. 

H., Nakahata, T. (2005). Prospective characterization of neural stem cells by flow cytometry analysis using a combination of surface markers. J Neurosci Res. 80, 456-66. 

After 48 h culture, collect cells into 4 mL FACS buffer. Take 300 pi cells to do the flow cytometry analysis for percentage of GFP-expressing cells. Normally, the good retroviral supernatant means the GFP% can reach to 90-95% at the 1 2 dilution, 75-85% at 1 8 dilution, and 60-70% at 1 16 dilution. 

Resuspend the cell pellet in 300 pi of FACS buffer and then perform flow cytometry analysis. 

In 2005, Lopez de Cerain et al. found that by using comet assays and flow cytometry analysis, compound 43 (Table 6) damages DNA under hypoxia and normoxia conditions [150]. In another study, it has been demonstrated that compound 43 produces reactive oxygen species (ROS) and oxidative DNA damage both in normoxic and hypoxic conditions into Caco-2 cells [151]. In normoxia, a significant increase of ROS was evidenced in a manner dose-dependent, while in hypoxia high ROS levels were observed at all the studied concentrations. Furthermore, modified comet assay demonstrated that oxidative DNA damage in normoxia and hypoxia was promoted by compound 43. 

So, these results showed a double effect of BOA on lettuce meristems an increasing significant delay in the cell cycle progression and a decrease in the mitotic index. Flow cytometry analysis showed a weak effect of BOA at cell cycle level in the step from G2 to M. BOA effect appeared to retard the cell cycle progression of treated-root meristems, which was very clear after 10 h BOA exposition (Fig. 12.5), and as an inhibition of the number of cells undergoing cell division. 

The effects exhibited in the flow cytometry analysis are also clearly detected by the mitotic index technique (unpublished data), which appears to be an excellent complementary technique in the cell cycle studies using flow cytometry. The mitotic index (see Fig. 12.5) revealed that the cell cycle progression goes slower in BOA-treated meristems than in control meristems and also that the maximum number of cells undergoing cell division is fewer and later in treated meristems (18% at 6 h BOA exposure time) than in control meristems (36% at 4 h distilled water exposure time). 

Then, and every 2 h, samples of BOA-treated plants and the corresponding controls were simultaneously processed using flow cytometry analysis. The 1 mm-apical tips of root meristems from forty BOA-treated plants were chopped with a razor blade in Galbraith buffer, supplemented with 100% Tween 20 and 100% beta-mercaptoethanol. Once filtered, the suspension, 5 pL of 1% RNase solution and 30 pL of 10 mg/mL ethidium bromide (EtBr), were added to 500 pi of filtered nuclei suspension and incubated for 30 min at room temperature in the dark. 

Re-suspend the resulting cell pellets in 250 pL ice-cold PBS containing 1% BSA. Keep the samples on ice until flow cytometry analysis. 

Figure 9.3 Mean total lymphocyte count after one course of treatment. Peripheral blood was collected from individuals administered test article (circles) or placebo (squares) on a weekly basis, and subject to flow cytometry analysis to determine mean lymphocyte counts. A standard panel of fluorochrome-conjugated antibodies was used to identify the various lymphocyte sub populations. The solid bar indicates the dosing interval.

Quantitative flow cytometry analysis, 430 Quantum confinement effect, 433,434, 436,445... 

The flow cytometry analysis proved that certain a-gal polymers were effective in inhibiting anti-Gal antibodies in human serum binding to pig kidney cells. Thus, such carbohydrate polymers can serve both as tools for evaluation of a-gal-anti-Gal interactions and as immimosuppressors in cell xenotransplantations. 

Srienc, F., Campbell, J. L., and Bailey, J. E. (1986) Flow cytometry analysis of recombinant Saccharomyces cerevisiae populations. Cytometry 7(2), 132-141. 

G3. Garovoy, M. R., Rheinschmidt, M. A., Bigos, M., Perkins, H., Colombe, B., Feduska, N., and Salvatierra, O. Flow cytometry analysis A high technology crossmatch technique facilitating transplantation. Transplant. Proc. 15, 1939-1944 (1983). 

Flow Cytometry Analysis. Flow cytometry analysis of blood and tissues has the advantage of allowing enumeration of lymphocyte subsets (see Chapters... 

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