setting up 6 color sort; questions

Dear all,

I would like to ask a question concerning 6 color sort: a method that we want to set up
in our lab. We want to look at hematopoietic stem cells (Lin-Sca+Kit+) and stain for an
additional three stemcell markers (all 3 rather low expressed.

We have in our lab the DIVA with detectors for
-FITC (530 +/-15)
-PE (585 +/-21)
-PE-TxR (630 +/-11)
-PE-Cy5 (675 +/-20)
-APC (660 +/-10)
-APC-Cy7 (710 Lp)
- and on the UV 424 +/-15

Also, we could get acces to an Aria that should have detectors for
FITC/PE/PE-TxR/PE-Cy5/PE-Cy7/APC/APC-Cy7 (+ Alexa Fluor. 430 and Pac.blue on the violet
laser). I would highly appreciate some feedback or tips on good color combinations to
choose. 

Is it correct that we should avoid low-expressing markers on APC, since one looses
low-APC-positive cells due to leakage-compensation in multicolorstainings?

Looking forward to you feedback. Thanks in advance!
Kees-Jan Pronk
_______________________________________________
- M.D., Dep. of Pediatrics, Lund University Hospital
- PhD-student, Center for Stemcellbiology and Cell Therapy, Lund University,
www.medfak.lu.se/stemcellcenter