Re: mouse eyes

From:Tracy Bergeron <TBergeron@criver.com>, TBergeron@criver.com


HI,

     I do mouse eyes on a regular basis.   We do not use any special
fixation methods that I have seen done with larger eyes (such as rabbit,
dog etc)..  The eyes are fixed whole in 10% NBF.
     Trimming:  I usually process the eyes whole, and have had no problems
doing so.
     Processing:    Our schedule of processing includes 1 station of NBF(20
min), 2 stations of 80% Flex(20 min each), 2 stations of 95% flex(20 min
each), 2 stations of 100% flex(25 min each), 3 stations of Xylene(20 min
each), and 4 changes of Paraplast x-tra(10 min each)..  This is our
standard mouse tissue program no special program for running mouse eyes.
     As for cutting.  I prefer to face the block off and allow it float in
my waterbath for a few seconds(bath set at 45 degrees C). Then I place the
faced warmed block into a  5% solution of fabric softener.   I cannot tell
you any specific amount of time to soak in the softener I actually have not
kept track of that timing..   The fabric softener seems to do wonders for
softening the lens enough to get nice unshattered sections.     Due to the
fact that this lab does not have a freezer to keep ice blocks or cold
plates I use a cryo spray to chill the block when I am ready to cut.  I
have found that Paraplast cuts best when it is quite cold.

     Hope this makes sense.

Tracy E. Bergeron
Histologist
Charles River Laboratories
Wilmington, MA
978-658-6000
x-1229









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