RE: Basic Storage of Blood

From: Olindo Assis Martins Filho (oamfilho@cpqrr.fiocruz.br)
Date: Thu Feb 10 2000 - 10:03:14 EST


Hi, Lee Anne Talbot

We have worke with dog lekocytes for flow cytometric analysis of CD markers 
using antibodies from SEROTEC. We have developed a protocol that we can 
lyse the whole blood using comercial FLS from BD and then kept the washed 
cells "fixed by Lysing solution" sterile in PBS 10% of FBS for over a week 
at 4 degrees. We did not find any increase on the autofluorescence for 
lymphocytes. We found out that CD14 for monocytes are lost after the 
fixation process and unfortunately if we need to quantify them we need to 
label cells right the way. However, for CD5, Thy, CD4, CD8, CD45RA, CD45RB, 
HLA-DR and CD21 no problems were detect comparing results before and after 
fixation.
If you need any aditional information I will be glad to help.
Unfortunately I do not have experience with rat cells.
Sincerely yours,


Olindo Assis
Centro de Pesquisas Rene Rachou - FIOCRUZ/BH
Brazil



This archive was generated by hypermail 2b29 : Sat Mar 10 2001 - 19:31:06 EST