Re: Slide "Plains"

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From:"Aidan C. Schurr" <> (by way of histonet)
To:histonet <>
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Just a couple of suggestions - the different planes you describe are quite
possibly due to different thicknesses of the mountant under the coverslip.
Too much mountant leads to a different refractive index, which can give the
problem you describe.  Not too sure with the haziness - could be the same
problem if the mountant layer is **very** thick, but this seems unlikely.
Another possibilty is that you have water in your zylene, or dehydration
has been incomplete.  You could perhaps try using two changes of xylene.

Hope this is of some help

Good luck
Aidan, New Zealand

>Hello Histonetters, I was hoping you could help me a couple problems.My
>pathologist was complaining that the stained H & E sections in our daily work
>was somewhat on different "plains" and needed constant focusing while under
>the scope. I sat down with her and went over some slides, these sections were
>somewhat "fuzzy or hazy" in some places and crisp in other, which makes me
>think possibly it's the mounting medium...we've been using this medium for
>sometime (resinous) and unfortunately we're on contract so my options are
>limited (for purchasing that is). Could it possibly be the deparaffination
>process? We microwave our sections to melt the paraffin, and put them in
>Xylene for 6 minutes to clear before sending them down the stainer. It does
>not happen on all the tissue, and there does not seem to be any preference as
>to type of specimen either (happens to bx's and surgicals). Any suggestions?
>Also, we seem to be getting holes in our blocks after embedding, the VERY
>irritating experience is becoming more and more frequent. We have fresh
>paraffin in our dispenser (and processor) and have a cold plate at optimal
>temperature, yet it seems at least 4-5 blocks per day are "breaking" when
>trimming because of these holes. Is it possible that we may be embedding too
>fast? Not giving the warm paraffin time to combine with the cold paraffin
>after inserting the tissue in the mold? Could it be that we aren't cleaning
>the molds enough? Again, any suggestions? Thanks for your help in advance, man
>I love this net......
>Kari Zajic HT,MLT
>Lead Histotech

	Aidan C. Schurr  BMedLabSc
	   Histology Service Unit		++64 3 4797152
	      Pathology Department
	         Dunedin School of Medicine


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