Flow cytometry evaluate

Figure 17.9 Rat in vivo hematotoxicity evaluated using flow cytometry (based on Saad et al. [59]). Gating strategy of flow cytometry evaluated rat bone marrow samples. N = at least 10000 cells. Results include absolute number nucleated cells, myeloid cells, nucleated erythroid cells and lymphoid cells.

The fluorochrome-conjugated monoclonal antibodies used may be varied. Investigation on one, a combination or all may be performed. In addition RNA Isolation to confirm the quality of RNA and Genochip Microarray for transcript profiling may be performed after flow cytometry evaluation. 

Evenson. D. P. (1989). Flow cytometry evaluation of male germ cells. In Flow Cytometty Advanced Re.tearch and Clinical Applications (A. Yen, Ed.), pp, 218—246. CRC Press. Boca Raton, FL. 

Akagi et al. demonstrated the use of nanoparticles composed of amphiphilic poly (amino acid) derivatives as vaccine delivery and adjuvants [62, 102-104]. To evaluate the uptake of OVA encapsulated within y-PGA-Phe nanoparticles (OVA-NPs) by DCs, murine bone marrow-derived DCs were incubated with 250 nm-sized OVA-NPs for 30 min at 37 °C. The cells were then analyzed by flow cytometry (FCM) and confocal laser scanning microscopy (CLSM). OVA-NPs were efficiently taken up into DCs, whereas the uptake of OVA alone was barely detectable at the same concentration of OVA (Fig. 13). OVA-NPs were more efficiently taken up than OVA alone by DCs, and the uptake of OVA-NPs was inhibited at 4 °C. These results suggest that OVA-NPs were phagocytosed mainly via endocytosis by the DCs. In the case of OVA alone, an approximately 30-fold... 

Ladies, G.S., et al., Phase two of an interlaboratory evaluation of the quantification of rat splenic lymphocyte subtypes using immunofluorescent staining and flow cytometry, Toxicol. Methods, 8, 87, 1998. 

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

FUNCTIONAL FLOW CYTOMETRY FOR EVALUATION OF IMMUNOTOXICITY AND MECHANISM OF ACTION... 

In summary, flow cytometry is clearly useful in evaluating macrophages and their role in toxicity. A major advantage of this technology is the rapid and accurate identification of subpopulations of responding cells from within a mixed population. There is no doubt that the utility of flow cytometry will increase in the future as new fluorescence probes are developed that allow investigators to more clearly assess various macrophage characteristics and the response of these cells to xenobiotics. 

Finally, multiparameter flow cytometry has the potential to improve existing methodologies for the evaluation of immunotoxicological endpoints such as the natural killer cell (NK) assay and the local lymph node assay (LLNA) to evaluate innate immunity and chemical-induced allergic disease. Accepted methodologies typically utilize radio-... 

De Guise, S. et al., Immune functions in beluga whales (Delphinapterus leucas) Evaluation of phagocytosis and respiratory burst with peripheral blood leukocytes using flow cytometry, Vet. Immunol. Immunopathol., 47, 351, 1995. 

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. 

These MHG class I alterations have been evaluated by the analysis of the panel of tumor samples with immunohistochemistry or flow cytometry in disrupted tumor cell suspensions as well as established tumor cell lines using a series of mAbs directed against HLA class I monomorphic, HLA-A or -B locus-specific, HLA-allelic epitopes or against various APM components including TAPI, TAP2, tpn and the different LMP subunits. The results demonstrated that rates of HLA class I and APM component losses in tumor cell lines strongly varied between the different tumor types analyzed. 

Jung, Y. S., Frank, J. F., and Brackett, R. E. (2003a). Evaluation of antibodies for immuno-magnetic separation combined with flow cytometry detection of Listeria monocytogenes.. Food Prot. 66,1283-1287. 

Morphometric and stereological research methods are valuable quantitative tools for toxicological studies. However, these methods can be very labour intensive and therefore are infrequently used for toxicity evaluation studies. Computerized image analysis may solve some of these problems in the fiiture. Other automated quantitative methods, such as flow cytometry, have also been used (Kangasniemi et al., 1990 Toppari et al., 1990). However, these are not routine techniques and are not incorporated in any testing guidelines at present. 

Telford, W G, King, L E, and Fraker, P J (1991) Evaluation of glucocorticoid-induced DNA fragmentation m mouse thymocytes by flow cytometry Cell Proliferation 24,447-459. 

Mysore, K. S. and Baird, V. (1997). Nuclear DNA content in species of Eleusine (Gramineae) A critical re-evaluation using laser flow cytometry. Plant System. Evol. 207,1-11. 

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