Hello Innocent,In my opinion, 12 hours in buffered formalin (pH 7.2) is not enough fixation time. Formalin idealy requires 40 hours to properly fix tissue. You may want to try a different fixative for lymph nodes.For processing, I would eliminate the 90% alcohol change and add a third absolute alcohol change. If you have space on your processor, add a third xylene as well. Sixteen hours sounds like a long time for processing. If some of that time is used for fixation, that is to your advantage. The length of processing really depends on the tissue. If you are processing lymph node only, then I recommend you lessen your times in each solution. Good luck,
From: Innocent Mosha [mailto:firstname.lastname@example.org]
Sent: Tuesday, June 17, 2003 2:16 PM
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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