Re: [Histonet] Let's Play solve this problem....

From:"Jackie M O'Connor"

Tim -   Are there any prizes for playing?

For IHC it would automatically set off incomplete fixation alarms.  How 
big are your tissues? What kind of tissue are we talking about?
If your tissues are not completely fixed in formalin (or whatever you fix 
in) they are getting primarily fixed in alcohol.
My second instinct would be contamination of your deparaffinization xylene 
(or clearing agent) in your staining set up, not allowing the hematoxylin 
to get to the nuclei because there is an invisible coating of paraffin - 
which of course is then completely removed in the last clearing xylenes.
Third - Incomplete processing - smell your block - no really, go ahead - 
smell your block.  Does it smell like any reagent in particular?  You 
shouldn't be able to smell anything if the processing was right.
But, I sense myself rambling . . . . 
Jackie O'

Timothy Macatee 
Sent by:
09/23/2005 09:49 AM

        To:     Histonetters 
        Subject:        [Histonet] Let's Play solve this problem....

Histonetters Assemble!

I'm having a recurring problem with my H & E stained slides.  The staining
is weak, especially away from the edges of the tissue.  It looks almost 
there is still an opaque paraffin haze to the tissue.  It could be a
processing problem because it occurs more towards the middle of the 
Is something not getting into the tissue far enough?  Is it Fixative (PFA 
Formalin), or processing reagents?  Or am I just whistling dixie?

Tim Macatee
NYU Medical Center
Research Histology Core

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