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!
Innocent
Postgraduate student,
University of Dar es Salaam
Muhimbili Campus.
Tanzania East Africa.
Fred Underwood
<funderwood@mcohio.org> 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.
Good
Luck,
Fred
-----Original Message-----
From: Innocent Mosha
Sent: Tuesday, June 17, 2003 5:16 PM
To:
Subject: How Can I Made Thin Sections For
H&E From Lymphnode tissue?
Dear fellows!
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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