Re: Bouins Fixation


I have done a lot of GMA work and when I used Bouins I had this problem.  It is
my opinion the the Bouins prevents infiltration and polymerization of GMA, I
come by this opinion by experience.  There are things you can do now to try and
recover the sample.  Face the block exposing the center tissue.  Take the whole
block on the block holder or remove the tissue from the block holder and
reinfiltrate using a shaker or ossilater/stirrer in GMA.  Reinfiltrate as long
as you can, then reembedd.  Polymizeration should be slow allowing the
initiater to penetrate to center of sample.  Another trick I use is to dry the
sample after it is reinfiltrated with a paper towel or tissue.  Lay the block
in a fume hood so that the exposed tissue is facing up, and drop on a drop or
two of the polymerization initiator right on the exposed tissue and let it sit
and penetrate for 5-10 min., wipe off excess initiator and reembed the sample
slowly, you can slow down polymerization by placing your sample in the fridge,
freezer or ice bath.  This will allow polymerization to reach the center of the
sample.  Sometimes this may only polymerize a thin layer of tissue, but it may
be enough to get the sections you need.  This process can be repeated as you
cut through the block.  In order to avoid the GMA infiltration inhibition of
Bouins in the beginning, wash the sample well in running dh20 after fixation
and before dehydration and infiltration of GMA.
Patsy Ruegg

louise renton wrote:

> Dear All,
> I need some advice please.
> We are currently using Bouin's fixative to fix rat tissue samples of muscle
> and bone. The samples are approximately 1cm x 0.5cm and were fixed for 5
> days prior to infiltration with Historesin (GMA). When cut, however, the
> centres were soggy and uncutable. Is this a problem with fixation or with
> the infiltration time being insufficient or both?. I seem to remember that
> way back when when I was learning about such, that Bouins has poor
> penetration, but excellent cytological preservation. What are the comments
> and thoughts of the forum?
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