[Histonet] Cuting speed and sem-thin plastic sectioning

From:Lawrence D Lanberg/O/VCU


   I  am a beginning microtomist - = I do this in between classes for the
   most part - in a Biochemistry laborato= ry. I semi-thin section murine
   retinas  using  plastic  (Spurr's  Formula).&nbs=  p;I  use  a Sorvall
   MT-1000  ultramicrotome.  My sectioning, staining and = destaining has
   developed  very  well  in  the  4 months I have done this. I have= one
   little  thing,  though,  that  seems to be impeding progress: Speed of
   cut= ting.

   Currently,  the only way I get quality sec= tions is when I cut at 0.3
   mm/sec.  Which is almost the slowest setting on t= he machine. If I am
   lucky,  I  may  get  three blocks done per hour; 20 section= s per-two
   slides per block. This is slow output, from what I am told.


   I  have experimented cutting these on a speed as high = as 7.3 mm/sec.
   This   greatly   increases my   output,   obviously,   but   after      staining-destaining  the  sections  appear  corrugated.  But  they are
   beautiful = at 0.3 mm/sec - the terribly slow speed.

   I  am  un= clear as to why the higher speeds I use give this unwanted,
   cordurouy-like  =  appearance  after  staining. Considering that I use
   plastic  (hard  formulation=  to  avoid  wrinkles) and that I cut with
   self-made  glass  knives,  can  anybody= provide a tip as to how I can
   possibly  cut  on  the  higher  speeds,  while  mai= ntaing quality of

   Thank You

   &= nbsp;

   Larry Lanberg

   Student Worker IST

   Dept. = of Biochemistry

   Virginia Commonwealth University

   (804) = 828-7974

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