RE: [Histonet] mordanting in Bouin's, storage in 70%, processing mouse tissues
Mallory's triple stain (I really had to dig into my histopast from
1962!) The reason mercuric chloride is used as a mordant for this stain is
that Zenkers (which contains mercuric chloride) was always a fixative of
choice for this special stain. One could actually fix in NBF, and mordant
in Zenkers before staining , when we did this stain in the first
I do not recall ever using Bouins as a mordant for the Triple, but John
Kiernan may set us straight on this whole subject. We only used Bouins
either as a fixative or mordant for Massons Trichrome, but if it works for
Mallory's, I would do it instead of toxic mercuric chloride.
Overprocessing of your mouse tissue sounds like the culprit for your dry,
brittle tissues. Mouse tissues are generally very small and can be
processed 30 to 45 min per change. Liver and brain must be totally fixed
before processing due to their homogenous nature or processing finished the
fixation and generally dries out the tissues even more.
70%, 80%, 95% x 2, 100% x 2, xylene X 2 with 3 changes of paraffin at 30
min each as heat will dry the tissues out even more.
After trimming a block, set it on cold ice water block, but return it to
the microtome and DO NOT TRIM OFF WHAT YOU HAVE JUST SOAKED!!!! You want
the first sections that come off the sharp blade. If your room is really
cold, you can even use a RT water soak, or try a gauze soaked in water over
the just trimmed block.
We simply wash Bouins fixed or mordanted sections in running tap water
until the color is gone, approx 5 min or so.
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-4303 (FAX)
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