Hi, Olindo Assis and anyone else working with canine leucocytes. We would be very glad to hear from anyone who is successfully applying markers to canine tissue that has previously been formalin fixed and paraffin embedded. Particularly pan lymphocyte or macrophage markers. Whose antibodies are you using and what protocols? Thanks Christina McCowan Veterinary Clinical Centre University of Melbourne At 13:03 10-02-2000 -0200, you wrote: > >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 >
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