Histotechnology Technical methods

STAIN FOR NEUROFIBRILLARY CHANGES IN CNS

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GALLYAS TECHNIQUE

SECTIONS

8µm 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

  1. Take sections to distilled water.
  2. Place in 5% periodic acid for 5 minutes.
  3. Wash in distilled water for 5 minutes twice.
  4. Place in alkaline silver iodide solution for 1 minute.
  5. Wash in 0.5% acetic acid for 10 minutes.
  6. Place in developer solution (prepare immediately before use) for 5-30 minutes.
  7. Wash in 0.5% acetic acid for 3 minutes.
  8. Wash in distilled water for 5 minutes.
  9. Place in 0.1% gold chloride for 5 minutes.
  10. Rinse in distilled water.
  11. Place in 1% sodium thiosulphate solution for 5 minutes.
  12. Wash in tap water.
  13. Counterstain in 0.1% nuclear fast red for 2 minutes.
  14. Wash in tap water.
  15. 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

NOTE: No responsibility is assumed by The University of Nottingham or the Queens Medical Centre NHS Trust for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions or ideas contained in the material herein. It is the users responsibilty to ensure that all procedures are carried out according to appropriate Health and Safety requirements.

© Copyright 1997 University of Nottingham Medical School Division of Histopathology. This page was last built on Mon, May 5, 1997 with Frontier. Thanks for looking in. Comments to James.Lowe@nottingham.ac.uk