Re: [Histonet] paraffin sectioning troubleshooting

From:Rene J Buesa

William:
  "Venetian Blind" effect is usualy caused by:
  1-blade holder or object too loose
  2-too steep blade angle
  3-tissue or paraffin too hard
  4-calcifiued areas in the block
  5-variations in the speed of sectioning
  6-too big block or
  7-unevenly har tissue
  The remedies could be, depending on the cause:
  1-tighten all movable parts holding blade holder and object
  2-reduce the blade angle
  3-use "heavy"duty disposable blades
  4-rehydrate and decalcify tissue
  5-try to cut with "even" strokes
  6-if the block is rectangular in shape, set the longer axis parallel to the blade's edge
  7-set the hardest areas to be cut first.
   
  If sections rolls over themselves this is usually caused by a too thick setting of the "thickness" knob = abobe 10 Ám. Poor infiltration or a hot block can be also the causes.
  Set your microtome to section at 5 Ám and cool the block adequately.
  Hope this will help you!
  RenÚ J.

William Ares  wrote:
  As a newbie to paraffin sectioning on a paraffin microtome, I've been plagued with differential results since starting. My current issue is that as they're cut, my sections are rolling back up on themselves instead of coming off the blade with the appearance of a venetian blind (as they were when the sections were looking good). As they appear brittle, I've tried putting the blocks in a humid environment to try to rehydrate them a little, but to no avail. Anyone have any good troubleshooting tips for me? Thanks in advance.

William Ares
Research Technician
Laboratory of Molecular and Developmental Neuroscience
Weill Medical College of Cornell University
Tel: (212) 746 5056
Email: wia2005@med.cornell.edu


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