http://www.nottingham.ac.uk/pathology/protocols/gallyas.html

SECTIONS


8m paraffin wax sections



SOLUTIONS


1. 5% Periodic Acid



2. Alkaline silver iodide solution



Sodium Hydroxide 40g

Potassium Iodide 100g

Distilled water 500ml
1% Silver nitrate 35ml

Dissolve the sodium hydroxide in water then add the potassium iodide and wait till dissolved. Slowly add the silver nitrate and stir vigorously till clear. Then add distilled water to give a final volume of 1000ml.


3. 0.5% Acetic Acid



4. Developer Working Solution



Add 3 volumes of stock solution II. to 10 volumes of stock solution I. stir and add 7 volumes of stock solution III. stir and wait to clear.



STOCK SOLUTION I



Sodium carbonate (Anhydrous) 50g

Distilled water 1000ml

STOCK SOLUTION II



Ammonium nitrate 2g

Silver nitrate 2g

Tungstosilicic acid 10g

Distilled water 1000g

Important: dissolve consecutively.



STOCK SOLUTION III



Ammonium nitrate 2g

Silver nitrate 2g

Tungstosilicic acid 10g

Formaldehyde (neat) 7.3ml

Distilled water 1000ml

Important: dissolve consecutively.


5. 0.1% Gold Chloride



6. 1% Sodium thiosulphate ("hypo")

7. 0.1% nuclear fast red 2.5% aluminium sulphate


METHOD



Take sections to distilled water.

Place in 5% periodic acid for 5 minutes.

Wash in distilled water for 5 minutes twice.

Place in alkaline silver iodide solution for 1 minute.

Wash in 0.5% acetic acid for 10 minutes.

Place in developer solution (prepare immediately before use) for 5-30 minutes.

Wash in 0.5% acetic acid for 3 minutes.

Wash in distilled water for 5 minutes.

Place in 0.1% gold chloride for 5 minutes.

Rinse in distilled water.

Place in 1% sodium thiosulphate solution for 5 minutes.

Wash in tap water.

Counterstain in 0.1% nuclear fast red for 2 minutes.

Wash in tap water.

Dehydrate, clear . Mount sections in DPX
RESULTS


--Neurofibrillary tangles, plaques neurites, and neuropil threads - Black


--Inclusions of multiple system atrophy - black


--Glial coils, threads and astrocytic plaques - black


--Nuclei - Red