RE: How Can I Made Thin Sections For H&E From Lymphnode tissue?
Thank you Fred!!
I agreed with you, that too much trimming of block and chill in Ice bath may give ice artefact on section as well shrinkage artefact. Over or underfixation could be the factor causing this problem, but why lymphnodes is the only victim? lymphoid tissue are rather solid, but it is possible to thin sections from cartilage, brain, liver even bones. A point of using new sharp knife is a temporary solution, especially for us poor resourse countries. With old sharpened knife we can make thin sections from various tissue, why not lymphnodes? Some of the helpful hints have used previously with little effect!
I think there might be confounding factors, other than what we have been discussing!!!!
Histonetters! do you occasionally face the similar problem? In need to here much from you!
University of Dar es Salaam Muhimbili Campus.
Tanzania East Africa.
Fred Underwood <email@example.com> wrote:
I worked with a pathologist who was a stickler about lymph node sections. He insisted on virtually all nodes being fixed in B5. Your processing schedule seems OK. I'll throw out a couple of helpful tips.
1.) If the block is crowded with nodes, divide it into one or two more blocks.
2.) Be gentle when facing the block. Cutting away too thick amounts will create a "moth eaten" effect microscopically.
3.) A new blade is a must. Although the edge may be too fine. Condition the blade by cutting sections from a plain block of paraffin.
4.) Do not chill the block with freeze spray. This will cause the tissue to crack within the block.
5.) If the nodes are fixed in B5, it is imperative to completely dry the section of all water before exposing to heat.
6.) If the nodes are not brittle, cool them without exposure to water. They can become somewhat spongy.
From: Innocent Mosha
Sent: Tuesday, June 17, 2003 5:16 PM
Subject: How Can I Made Thin Sections For H&E From Lymphnode tissue?
I`m really experiencing difficulties in making thin section (5microns) for H&E from lymphnode tissue. Lymphnode tissue are usually prompt/immediately fixed in buffered formalin (pH 7.2) for 12 hours before runned in a automated machine for 16 hours. In this machine tissue goes through and agitated in different baths; starting with 70% alcohol (two changes), then 90%, 95% and absolute alcohols (all these in two changes). Then two changes of Xylene follows before embeded in paraffin wax.
The quality of H&E sections of other tissue is good, but with lymphnode the section are thick/poor quality and impose difficulties in making diagnosis.
I appreciate your opinion/advice, so as I can
improve the quality of sections for well fare of patients in this part of the world.
Thanking you in advance.
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