RE: Hirschprung's for J.A. Kiernan

From:"J. A. Kiernan" <>

On Thu, 12 Apr 2001, Stephanie Moore wrote:

> I noticed that your newest book has an updated AChE protocol.  I have
> ordered this book through our university library, however it has not yet
> arrived.  Before I try this, I just want to verify something.

The new publisher (Arnold) assures me that the 2000 reprint of 
the 3rd edition is now available in N. America, where it's
distributed by Oxford Univ. Press. I still get lots of emails
from people who say they can't get it. 

> Summary questions:
> Should I use the diluted medium for my brain sections?
> Which enhancement method works best?

I'd recommend trying both the stronger and the more dilute
medium, then settle for whichever is better. OsO4 isn't a
good way to enhance DAB for LM, especially of frozen
sections, because it darkens the tissue in its own right
(mainly by being reduced by unsaturated bonds of lipids).
Inclusion of Ni or Co is, as you point out, much more 
sensible. For really strong activity (motor end plates,
or striatum in the brain), you don't need the DAB
enhancement because the copper ferrocyanide reaction
product is a good strong brown.

Have a good Easter weekend.
                             John Kiernan
John A. Kiernan
Department of Anatomy & Cell Biology
The University of Western Ontario
London,  Canada   N6A 5C1

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