Re: cytotoxicity testing

From: Mark A. KuKuruga (kukuru@umich.edu)
Date: Fri May 19 2000 - 07:43:57 EST


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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