Abstract 4054: Cell viability, apoptosis and ploidy analyses on a multi-laser, versatile flow cytometry analyzer – the EclipseTM

Abstract

Abstract Cell viability, apoptosis and ploidy analyses are essential tools in cancer research involving in vitro and in vivo studies. Assays for cell count and viability, apoptosis and cell cycle analysis are described in this report. Jurkat or Chinese hamster ovary cells were treated with various reagents such as anti-CD95, taxol, or brefaldin A to induce apoptosis. Apoptotic cells were labeled with Avenxin-V conjugated with Pacific Blue or FITC to determine externalization of phosphatidylserine to the cell surface. In combination with cell impermeant dyes, such as 7-aminoactinomycin D (7-AAD) or propidium iodide (PI), live, early apoptotic and late apoptotic/dead cells were labeled and analyzed on an Eclipse equipped with the 405 and 488 lasers. The iCyt® Eclipse TM is a compact bench top flow cytometry analyzer with an autoloader that supports a wide variety of multi-well plate and test tube formats. The system can be equipped with up to 4 lasers (405, 488, 561 and 642nm) and provides simultaneous detection of 5 colors, forward scatter, side scatter, electronic volume (EV) and cell concentration. The EV parameter enables accurate cell volume (size) measurement and allows normalization of fluorescent intensity on a per volume or surface area basis. Apoptotic cells were also detected with a phycoerythrin-labeled anti-active caspase 3 antibody after fixation and permeabilization. Cell cycle analysis with G0/G1, S and G2/M phase statistics plus cell size was achieved by labeling cells with 4′,6-diamidino-2-phenylindole (DAPI) in a cell permeabilization solution and analyzing on the Eclipse. Together with its intuitive software, the Eclipse flow cytometry analyzer provides a rapid, accurate and cost effective tool for most flow cytometry applications such as cell viability, apoptosis and cell cycle analysis. Note: For research use only. Not for use in diagnostic procedures. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4054. doi:10.1158/1538-7445.AM2011-4054