[Histonet] Paraffin Tissue Crumbles

Michael Mashore mmashore <@t> vapop.ucsd.edu
Tue Nov 9 16:56:49 CST 2010

Hello Histonet Users,


I have just started using paraffin and am having many difficulties. Most of
the time my tissue crumbles when sectioning. I have no real experience in
paraffin histology and have been given the task of becoming proficient by
myself, so I am hoping for feedback as to why my tissue keeps crumbling. The
tissue in question has been:  skeletal muscle, cardiac muscle, liver, and
brain (all from rat).


The tissue was fixed in 10% neutral buffered formalin for 7 days at 4°C and
then transferred to an automated tissue processor, with the following


2 hours 70% dehydration alcohol 

2 hours 80% dehydration alcohol

2 hours 95% dehydration alcohol

2.5 hours 95% dehydration alcohol

2 hours 100% dehydration alcohol

2 hours 100% dehydration alcohol

2 hours 100% dehydration alcohol

.5 hour Hemo-De

.5 hour Hemo-De

.5 hour Hemo-De

1 hour paraffin

4 hours paraffin


They were infiltrated for 1 hour without vacuum then embedded.

The blocks were stored in the freezer before cutting.

The knife angle was 5°. 

Sections were 5µm thick.


I would appreciate any feedback whatsoever. 


Thank you very much.



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