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