RE: Fix & Perm + CCA
kukuruga%kasle1.dnet.wayne.edu@rocdec.roc.wayne.edu
Wed, 31 Jan 1996 12:58:41 -0500
Try supplementing the Caltag reagent with a low concentration of detergent --
Triton X100, NP40, Tween20, etc. -- this will improve stainablilty with PI.
Be sure to start with an extremely low concentration, however, since your
cells will probably be more susceptible to the treatment. Check surface
marker fluorescence and light scatter to be sure these parameters are
adequately preserved and correlate well percent-wise with un-supplement-
treated cells.
Alternatively, you could use H33342, or DAPI. These staines enter PFA-fixed
cells quite readily, so work well with Fix/Perm reagents. These are UV
excitable, unfortunately, so you usually need a multi-laser machine.
Mark KuKuruga, Karmanos Cancer Inst
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
and distributed free of charge as an educational service to the cytometry community.
If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
