Histological Staining Techniques

Gordon and Sweets' method for reticulin
Gordon & Sweets, 1936.
Gelatine coated slides recommended.
  • Dewax sections, rinse in alcohol, rinse in water.
  • Oxidise in acid potassium permanganate - 5 minutes.
  • Wash briefly in water.
  • Bleach in 1% oxalic acid - 2 minutes.
  • Rinse in distilled water followed by a thorough wash in tap water.
  • Sensitise in 2.5% iron alum - 10 minutes.
  • Wash in several changes of distilled water.
  • Treat with silver solution in a coplin jar - 5 seconds.
  • Rinse briefly in distilled water.
  • Reduce in 10% 'Analar' formalin in distilled water - 1 minute. (In fume hood.)
  • Wash in tap water followed by distilled. (In fume hood.)
  • Tone in 0.2% gold chloride - 2 minutes.
  • Wash briefly in distilled water.
  • Fix in 5% sodium thiosulphate - 2 minutes.
  • Wash well in tap water.
  • Rinse in distilled water.
  • Counterstain with nuclear fast red - 5 minutes.
  • Wash in distilled water.
  • Dehydrate, clear and mount.

Results:

 
Reticulin - black.
Nuclei - red.
Collagen and cytoplasm - purple/pink.
 

Acid potassium permanganate
0.5% potassium permanganate in 3% sulphuric acid

Gold chloride = sodium tetrachloroaurate

Silver solution
To 5mls of 10% silver nitrate add strong ammonia drop by drop until the formed precipitate just redissolves.  Shake the solution for a few seconds after adding each drop.  Add 5mls 3% sodium hydroxide and redissolve the precipitate with a few more drops of ammonia.  Again, shake the solution after adding each drop.  The finished solution should have a slight opalescence.  If the solution goes completely clear add a drop or two of 10% silver nitrate until a slight opalescence remains.  Make up to 50mls with distilled water.  Kept in a sealed coplin jar at 4oC the solution can be used for up to one month.

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