M, You really shouldn't have any problem co-staining with a PE moab . . . you may have to play with concentrations of the viability/cytotoxicity indicators, to better accommodate the signal crossovers. Depending on the machine (whether or not you can change filters), using a narrower bandpass for the PE (I sometimes use a 575/14) can reduce compensation problems. If you can go APC, much better. MAK. MSchwa3722@aol.com wrote: > I'm wondering if anyone has used Molecular Probes reduced biohazard > viability/cytotoxicity test. I'm testing activated cells effect on a breast > cancer cell line in a series of T cell:Tumor cell dilutions in which the > tumor cell is greatly reduced in number. I performed a similar test > previously using a green lipophillic dye (DiO18) counterstained with PI. The > syto/dead red dyes of this kit have the advantage of being able to stain the > cells simultaneously. My question is what color combination should I use to > distinguish the tumor cells from the T cells. Syto and dead red flouresce in > FL1 and FL3 but I'm wondering, if anyone has used this kit before, has there > been any difficulty in compensating a PE conjugated antibody or should I > consider using an APC conjugated Ab? Thanks in advance for your help. -- Mark A. KuKuruga, Managing Director University of Michigan Core Flow Cytometry <http://www.cancer.med.umich.edu/flow_cytometry> phone: 734-647-3216 fax: 734-936-7376 kukuru@umich.edu
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