[Histonet] CD34 in trephine bone marrow

[Gudrun Lang]

Hi,
Short time ago I've asked for CD34 protocols. Thank you for the answers, but
I still have troubles.
Our treatment:
Trephine bone marrow biopsies - ca. 3-4 mm in diameter, 15-25 mm long
Fixation - 12 h 8% neutral buffered formaldehyd
Decal - 6-8 h "Cal-rite", 5-10% formic acid and formaldehyd, roomtemp.
Dehydration and infiltration in paraffin - VIP over night
Immunostainer - benchmark XT
CD34, Dako QBEND10 1:50

I've tried several modification with the protocols. In fact the tonsil on
the same slide shows positiv cells with every method, also in the bone
marrow biopsy there are positiv endothels (no retrieval, 24 min), but the
lymphoblasts are all negative. I think the antibody is allright, but
something has happend to the lymphoblasts due to the pretreatment.

Any suggestions are appreciated.

Thank you 

Gudrun Lang
Akh Linz
Austria