Llewellyns Double Oxidation, Thiosemicarbazide, Schmorl

Monday, November 24, 2003

Description
Staining protocol

Procedure
Bring sections to water via xylene and ethanol.
Do a Mallory bleach.
Wash with running tap water
Oxidise in solution A for 10-20 minutes.
Rinse well with water.
Place into solution B for 30 minutes.
Wash well with running tap water.
Return to solution A for a further 20 minutes.
Rinse well with water.
Place into solution C for 10 minutes.
Wash well with running tap water to remove all traces of thiosemicarbazide.
Place into freshly made solution D for 10 minutes.
Wash with running tap water.
Optionally, counterstain with nuclear fast red or lightly with eosin.
Rinse well with water.
Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected Results

Fungi - blue
Nuclei - red (if used)
Background - pale pink (if used)

Recipes
Solution A
1% aqueous periodic acid

Solution B
Aniline 10 mL
Glacial acetic acid 90 ml

Solution C
1% aqueous thiosemicarbazide

Solution D
1% aqueous ferric chloride 30 mL
1% aqueous potassium ferricyanide 4 ml
Distilled water 6 ml
This solution should be used fresh.
Add together immediately before using.

Fixation & Sections
Most fixatives should be satisfactory. Paraffin sections.

Supplies

Tips

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Llewellyns Double Oxidation, Thiosemicarbazide, Schmorl