There is no single Gallyas method.
Ferenc Gallyas published several silver staining methods for paraffin embedded tissues, especially CNS. They have in common: accurate control of silver and silver-diammine ion concentrations (by using ammonium nitrate and NaOH rather than ammonium hydroxide), and stabilization of the physical developer with tungstosilicic (= silicotungstic) acid. Specificity for such things as microglia, neurofibrillary tangles, capillaries and immunohistochemical reaction (DAB) products is determined by pre-treatments.
Don't do a Gallyas method on the strength of someone's typed worksheet or an internet posting. Get your instructions from a primary source. The following references may be helpful. This list does not include all Gallyas' papers, only the ones that I have read and found interesting enough to record.
Gallyas (1963) Silver microglia method. Acta Neuropathol. (Berl.) 3: 206-209.
Gallyas F (1979) Light insensitive physical developers. Stain Technol. 54: 173-176.
Gallyas F (1980) Chemical nature of the first products (nuclei) of the argyrophil staining. Acta Histochem. 67: 145-158.
Gallyas F, Gorcs T, Merchenthaler I (1982) High-grade intensification of the end-product of the diaminobenzidine reaction for peroxidase. J. Histochem. Cytochem. 30: 183-184.
Gallyas F, Wolff JR (1986) Metal-catalysed oxidation renders silver intensification selective. Applications for the histochemistry of diaminobenzidine and neurofibrillary changes. J. Histochem. Cytochem. 34: 1667-1672.
Gallyas F (1971) Silver staining of Alzheimer's neurofibrillary changes by means of physical development. Acta Morphol. Acad. Sci. Hung. 19: 1-8.
Gallyas F (1971) A principle for silver staining of tissue elements by physical development. Acta Morphol. Acad. Sci. Hung. 19: 57-71.
Merchenthaler I, Stankovics J, Gallyas F (1989) A highly sensitive one-step method for silver intensification of the nickel-diaminobenzidine endproduct of peroxidase reaction. J. Histochem. Cytochem. 37: 1563-1565.
Gallyas F, Guldner FH, Zoltay G, Wolff JR (1990) Golgi-like demonstration of 'dark' neurons with an argyrophil III method for experimental neuropathology. Acta Neuropathol. (Berl.) 79: 620-628.
Gallyas F, Zoltay G (1992) An immediate light microscopic response of neuronal somata, dendrites and axons to non-contusing concussive head injury in the rat. Acta Neuropathol. (Berl.) 83: 386-393.
Gallyas F, Zoltay G, Balas R (1992) An immediate light microscopic response of neuronal somata, dendrites and axons to contusing concussive head injury in the rat. Acta Neuropathol. (Berl.) 83: 394-401.
Gallyas F, Zoltay G, Horvath Z (1992) Light microscopic response of neuronal somata, dendrites and axons to post-mortem concussive head injury. Acta Neuropathol. (Berl.) 83: 499-503.
Gallyas F, Zoltay G, Dames W (1992) Formation of 'dark' (argyrophilic) neurons of various origin proceeds with a common mechanism of biophysical nature (a novel hypothesis). Acta Neuropathol. (Berl.) 83: 504-509.
Gallyas F, Hsu M, Buzsaki G (1993) 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.
Gallyas F (1982) Physicochemical mechanism of the argyrophil I reaction. Histochemistry 74: 393-407.
Gallyas F (1982) Physicochemical mechanism of the argyrophil III reaction. Histochemistry 74: 409-421.
Gallyas F, Merchenthaler I (1988) Copper-H2O2 oxidation strikingly improves silver intensification of the nickel-diaminobenzidine (Ni-DAB) end-product of the peroxidase reaction. J. Histochem. Cytochem. 36: 807-810.
Some of Gallyas' methods have been improved upon by others, including Braak and Munoz in the context of pathology of Alzheimer's disease and other dementias.
These and related methods were reviewed by Bill Grizzle (1996) Theory and practice of silver staining in histopathology. J. Histotechnol. 19: 183-195. This is in a special issue of the Journal of Histotechnology (Vol 19#3) devoted to silver staining. Read it. You may find a technically simpler but adequate alternative to the Gallyas method you were asking about.
Gallyas' techniques are for the "advanced" worker (technician or academic) who understands silver staining techniques. They won't work in the hands of a new graduate student or a molecular biology postdoc with no prior instruction or experience in using methods of this kind.
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----- Original Message -----
From: Sharon E Willman
Date: Monday, April 28, 2008 14:56
Subject: [Histonet] Gallyas Stain
> Does anyone have a staining protocol for paraffin embedded
> using the Gallyas method? I would appreciate any
> information you might
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