sectioning bubbles

Scott Douglas douglas at utsc.utoronto.ca
Thu Apr 4 19:46:35 EST 2002


I have a rather frustrating problem regarding the sectioning and
mounting of Paraplast Plus-embedded Arabidopsis tissue.  Following
sectioning and transfer of ribbons to Probe-on-Plus slides (Fisher), I
find that water bubbles form between the slide and the ribbon, often
resulting in puncturing of the paraffin.  A good deal of my sections are
being ruined by this phenomenon.  The methods I have used to transfer
the ribbon to the slide are as follows:
1. Floating the ribbon on a 42C water bath for a minute or so followed
by transfer to the slide.  Excess water is shaken off and the slide is
moved to a 42C slide warmer.
2. Placing the ribbon directly onto a slide with water already on it.
Slides are transferred to a slide warmer until ribbons have expanded and
excess water is removed using the corner of a paper towel.
In both cases, bubbles always develop under the ribbon upon incubation
at 42C.  I have tried using gelatin-coated slides and encounter the same
problem.  Furthermore, the wax I am using is brand new and its
temperature has never exceeded 62C.
If anyone has any suggestions as to how I could solve this problem I
would be very grateful.

Scott Douglas
douglas at utsc.utoronto.ca
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