CD45 etal in rabbit spleen

From:Kristin Gebhard Mitchell

A collegue brought me this dilemma and I'll pass it along to ya'll...

She has frozen (not well) rabbit spleen that was post fixed in Formalin. 
She used CD45 (don't know the clone) and Ki67 and some kind of 
macrophage ab (not CD68) as her primary abys (seperatly), the Vector 
ABC-HP kit and used AEC as her substrate. There was no pretreatment and 
no counter stain.

The CD45 has high background but the no primary control was completly 
clear. The others stained well...but the staining pattern is confusing.

The actual problem is that she's not certain of what it is SUPPOSED to 
look like  :-)

They were expecting clusters of positive cells (specific to the white 
pulp?) instead they got CD45 ALL OVER, Ki67 randomly and I never saw the 
macrophage but I have the impression it was all over.

I've searched thru the suggested IHC and staining links and various 
search engine for examples of spleen IHC, specific for these abys or for 
CD3 or some other lymphocyte marker to see what a normal pattern would 
be, but no luck. And the various histology books around here don't have 
a large selection of spleen material, let alone IHC on spleen.

Any ideas? Any ideas on what to search for? Any help is appreciated!

Thanks!  Kristin


Kristin Gebhard Mitchell
Histology Core Lab
Department of Ophthalmology
University of Minnesota
370 Lions Research Building
2001 6th St SE   Mpls, MN 55455

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