Karen:

As John Kiernan has mentioned, these methods by Gallyas* are difficult. They
are especially tricky to trouble-shoot on such thick sections. A
trouble-shooting chart is available on below reference.

If the student is attempting the Gallyas technique for neurofibrillary
changes in CNS, then the preparation of the stock solutions for the working
developer is critical. The order of the chemicals into the DH20, must be
dissolved thoroughly and consecutively.

If another technique is an option, I would also suggest the Luxol fast
blue/cresyl violet or the Luxol fast blue/PAS method (Kluver and Barbera).

*Gallyas, F. , Hsu, M. , and Buzsaki, G. Four modified silver methods for
thick sections of formaldehyde-fixed
     mammalian central nervous tissue: 'dark' neurons, perikarya of all
neurons, microglial cells and capillaries. J. Neurosci.
     Methods 50:159-164, 1993.

Eric Kellar
Histology/Immunohistochemistry
University of Pittsburgh Medical Center.
	----------
	From:  Karen D. Larison [SMTP:LARISONK@UONEURO.uoregon.edu]
	Sent:  Thursday, November 19, 1998 10:06 PM
	To:  HistoNet@Pathology.swmed.edu
	Subject:  myelin stain

	Histonetters,

	A graduate student here is attempting to stain 50 micron brain
sections for
	myelin using the Gallyas stain, but isn't having much luck with
stain
	penetration.  Is there anything he can add to his solutions to
increase stain
	penetration, or is there an alternate myelin stain he could use on
these thick
	sections?

	I am so greatful to this community of histologists for their
knowledge and
	their willingness to share their expertise.

	Karen Larison