Dead cell exclusion in FL4

Hi everyone,

1. We'd like to do some 4-5 color analysis with dead cell exclusion in
FL4. Anybody has any experience with TOPRO-3 and sorts (Mol. Probes)?
How does it compare (sensitivity/specificity) with PI or 7-AAD?

2. When doing dead cell exclusion, is it necessary to RNAse-treat cells
before applying the DNA-probe (like you would do for cell cycle
analysis); we're working with highly activated cells with high RNA
synthesis and we're concerned about possible uptake of the
nucleotide-probe by mRNA.

Thanks a lot!

Karim Vermaelen

Pulmonary Immunobiology
Ghent University Hospital
Belgium