Are you certain that it is formalin and not paraformaldehyde? The latter
will get cloudy. Also, is there any way that your clearing solvent might be
making its way in to the formalin solution?
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1) Acid alcohol: I use several drops of concentrated HCL in 95% ethanol (1%
HCL in 70% alcohol well too). Simply dip your rinsed slides (or sections)
into solution a couple of times, rinse and check them... a dip in water with
about 10drops of ammonia should follow as well...
2) Saturated picric
I don't know whether you are decalcifying tissue or not, but I have had
similar problems with tissue that was over-decalcified. It was particularly
bad when decalcified with commercial decalcifying agent which contained
formic along with several chelating agents. The end result was tissue that
I don't think the bone matrix will re-calcify with normal bone histo
solutions. What exactly are you using to decalcify the tissue? Hardening
can occur at many steps following decal. I stopped using a commercial
decalcifying agent (which consisted of formic acid with chelating agents
added)
I think that your problem has mostly to do with inadequate
dehydration/infiltration. I've had this problem mainly with sections of
vertebrate skin or sections that possess lots of little blood vessels.
Water is either unable to move through the keratin sufficiently to
dehydrate and I often
So, there are tons of hematoxylin recipes out there, and many, I find, are
very similar in their staining properties. Yes, Richard Allen manufactures
a modification of Harris' Hematoxylin, which is essentially Harris' recipe
without the mercuric oxide. I don't know how long Richard Allen's
I don't know that there's a way to change the pH of water through
filtration. The pH is affected mostly by dissolved gasses and ions which
aren't easily removed through filtration alone. You can adjust the pH by
using buffered solutions, but then its really not water anymore.
If you just have
I'd go for the old AO 820. They can be used by any and everyone and its
nearly impossible to break them.
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I think that distilled water starts out at a ph of around 6-6.5, but usually
DI water is closer to 7. Over time, both will drop in pH unless you buy
specialized housing for the DI water. Since many people doing IHC at my
university are using RO water, which has a pH similar to what the pH of the
I think that distilled water starts out at a ph of around 6-6.5, but usually
DI water is closer to 7. Over time, both will drop in pH unless you buy
specialized housing for the DI water. Since many people doing IHC at my
university are using RO water, which has a pH similar to what the pH of the
I think that distilled water starts out at a ph of around 6-6.5, but usually
DI water is closer to 7. Over time, both will drop in pH unless you buy
specialized housing for the DI water. Since many people doing IHC at my
university are using RO water, which has a pH similar to what the pH of the
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