GBL® ROSA-STAIN

5058-Grocott Methenamine Silver Staining Kit

Title Description
REF No 5058
Purpose It is used in the examination of fungi by light microscopy.
Ingredients A) Chromic acid solution: Potassium chromate
B) Sodium metabisulfite solution: Sodium metabisulfite
C) Methenamine solution: Methenamine
D) Silver nitrate solution: Silver nitrate
E) Borax solution: Sodium tetraborate
F) Gold chloride solution: Gold (III) chloride trihydrate
G) Sodium thiosulfate solution: Sodium thiosulfate
H) Light green solution: Light green SF yellowish, Glacial acetic acid
Working Principle Grocott Methenamine Silver Staining Kit is a histological staining method used for the detection of fungi and other microorganisms. It is used especially for the visualization of fungi such as Pneumocystis jirovecii as well as some bacteria and protozoa. The dye targets the cell walls or capsules of these microorganisms and reveals them as black or dark brown under the microscope. Chromic Acid; oxidizes tissues and causes polysaccharides (such as glycans) in the tissues to turn into aldehyde groups. This transformation creates sites where silver ions can bind. Methenamine Silver Solution; The combination of methenamine and silver nitrate reacts with aldehyde groups in the tissue. This reaction allows silver ions to bind to the cell walls or capsules of microorganisms. Gold Chloride; is used to make silver deposits more visible. Gold chloride increases the stability of silver and makes it appear with a darker color. Sodium Thiosulfate; removes excess silver ions, cleans the background and increases the clarity of the staining. Light green SF yellowish; stains the tissue background green, allowing microorganisms to be clearly seen as black or dark brown/grey.
Warnings, Precautions, Recommendations Caution. Danger. Obtain special instructions before use. May cause allergic skin reactions. Causes severe skin burns and eye damage. May damage fertility. May damage the unborn child. May cause genetic defects. Harmful if swallowed.
Decomposes toxic gases in contact with acids. May cause cancer by inhalation. Very toxic to aquatic life with long lasting effects. Avoid release to the environment. Wear protective gloves/protective clothing/eye protection/face protection. IF ON SKIN: Wash with plenty of soap and water. IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. IF exposed or concerned: Get medical advice/attention.
Application 1. Cover the preparation with “A- Chromic acid solution”. After 20 minutes, shake off the preparation and remove the reagent.
2. Wash with distilled water for 5 seconds.
3. Cover the preparation with “B- Sodium metabisulphite solution”. After 1 minute, shake off the preparation and remove the reagent.
4. Wash with distilled water for 5 minutes.
5. Immerse the preparation in “Methenamine silver solution”. Incubate at 60°C for 60 minutes.
6. Cool to room temperature for a maximum of 10 minutes.
7. Wash with distilled water for 5 seconds.
8. Cover the preparation with “F- Gold chloride solution”. After 3 minutes, shake off the preparation and remove the reagent.
9. Wash with distilled water for 10 seconds.
10. Cover the preparation with “G- Sodium thiosulfate solution”. After 5 minutes, shake the preparation to remove the reagent.
11. Wash with distilled water for 10 seconds.
12. Cover the preparation with “H- Light green solution”. After 30 seconds, shake the preparation to remove the reagent.
13. Wash with distilled water for 10 seconds.
14. Dehydrate, clean, cover with synthetic resin and examine under a microscope.
Theoretical Results
Mushrooms Black
Mucin Dark Grey
Backgrounds Green

Related Media
Shelf Life 1 year
Shelf Life After Opening the Lid
Storage Temperature
pH (at 25°C)
Transportation Information
Ordering and Packaging Information
Order No Unit Volume Packaging Type Quantity in a Box
445058050050 Piece 50 mL 50 ml, White Bottle with HDPE Dropper 1 kit

Keywords Grocott Methenamin stain, Mushrooms stain,
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