Grocott Stain Kit

Description

Kit Contains:
Chromic Acid 5% Solution x 2
Hexamine (Methanamine) 3% Solution x 2
Silver Nitrate 5% Solution
Sodium Tetraborate 5% Solution x 2
Sodium Metabisulphite 1% Solution
Gold Chloride 0.1% Solution
Sodium Thiosulphate 2% Solution
Light Green (0.2% in 0.2% Acetic Acid)

General Information:
Procedure Time : 105 minutes (post warming of working solution) (approximate)
Shelf Life: 3 Years (from date of manufacture)
Storage:    Silver Nitrate 2-8 ° C. Other reagents 15-25 ° C

Results: 
Fungi, pneumocystis, melanin, Basement Membrane:  Black
Mucins, Glycogen:                                                                  Beige/Grey
Background:                                                                            Pale Green

Working Stock Solution
Hexamine-Silver Working solution:
In a chemically clean container measure 23ml of hexamine and 1.25ml of 5% silver nitrate. A white
precipitate forms which will dissolve on shaking
Add 3ml of 5% sodium tetraborate and 25ml of distilled water. Make fresh every time.
Filter the solution into a Coplin jar and put into an oven or water bath at 56°C in darkness. Check
temperature of solution before starting

Protocol:  

1. Dewax sections, hydrate through alcohols and rinse in tap water. For smears flood slides with distilled water
2. Oxidise the sections with chromic acid for 60 minutes at room temperature
3. Rinse in distilled water
4. Treat sections with sodium metabisulphite solution for 1 minute
5. Wash well in running tap water and then in several changes of distilled water
6. Stain sections in the pre-heated working silver solution at 56°C for 15 to 20 mins until section turns yellowish-brown (check microscopically after washing in distilled water – fungi should be dark brown). If staining not complete return to the heated silver solution for 2-3 minutes and repeat the examination process
7. Wash in several changes of distilled water
8. Tone sections with gold chloride for 5 minutes
9. Wash in several changes of distilled water
10. Treat with sodium thiosulphate for 5 minutes
11. Wash in distilled water and counterstain in light green solution for 30 secs
12. Dehydrate, clear and mount

Hazard Information

UN3082