Re: Junks on Stained (Frozen Lymphoid) Tissue

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From:Barry Rittman <>
Date:Fri, 19 Mar 1999 07:25:15 -0800
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		most tissues which have a high ratio of cells to extracellular matrix
have a tendency to show cracks in sections, with both frozen an paraffin
sections. This is because there is not much keeping the cells in position.
You can therfore  end up with a situation where the cells or fragments of
them float off the section and become relocated, whether we are talking
about floating sections on a water bath or simply placing in staining
reagents. I found this to be especialy noticeable with spleen. I believe
that this is a possibility.
What happens if you cut thicker sections? If sections are say 6-7 microns
yu may be able to idntify specific the debris as specific cell fragments if
these are the problem.

Other posibilities are 
if you allow slides to dry excessively once cut or do not fix for an
adequate period of time
if the blocks that you are sectioning are drying in the cryostat then
similar tissue pieces may also be generated during sectioning,
if, as noted by previous writer the slides are dirty.
If you have squames on the sections these will stain a bright red in H and
E and will be roughly hexagonal in shape. 
hope this helps

At 07:16 PM 3/18/99 -0500, you wrote:
>	Frozen lymphoid tissues after sectioning and staining features some
>debris (hair like, glass like structures and other junks). It is very
>annoying when you have it sitting right at the center of a picture perfect
>follicle. Does any one have an idea of the source of these debris?  I think
>that this occurs, during sectioning but the source of these debris I have
>not been able to determine.
>	Again, thanks to all of you that gave suggestions on methods of
>freezing lymphoid tissues. I have recently made excellent sections (cracks
>and holes free) from a rabbit appendix tissue frozen in OCT placed in a
>mixture of isopentane and dry ice.
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