[Histonet] HELP! vertical lines in paraffin

From:dgaupp@tulane.edu

Histonetters:


I am having problems sectioning mouse ovary tumor without vertical lines from
the bottom of the block(paraffin only - no tissue touching the blade)  all the
way to top of block.  I did not process/embed these tissues.  So far I've done
the following:

       1.  changed blades(went threw about 10 just in a few blocks)
       2.  cleaned microtome
       3.  changed angles(6-12 degree & everything in between)
       4.  sectioned hot, cold, freezing
       5.  soaked blocks in ice cold water; froze blocks
       6.  surfaced decal 5 - 40 minute intervals
       7.  re-embedded
The paraffin seems to be causing the lines!  I've never experienced problems
like this before.  Usually its the tissue itself that causes a nick in blade
which therefore causing vertical lines to appear when sectiong the tissue.
Help is needed! What next is recommended?



Dina Gaupp, BS MT
Tulane University Medical Center
Center for Gene Therapy
Histology Core Facility
1430 Tulane Ave
New Orleans, La 70112
Lab phone 504.988.1194
email dgaupp@tulane.edu



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