ALKALINE METHYLENE BLUE SOLUTION
Methylene Blue Solution Citric Acid 0.1M Glycine Buffer (pH 10.6) 2 small, clean (100-125ml) Glass Bottles
Sterile Pipets Distilled Water Mechanical Pipeter Foil
1. Weigh 0.1g Methylene Blue and place in small glass bottle.
2. Add distilled water up to 100ml mark on bottle.
3. Swirl bottle to dissolve media. (This is your 0.1% Methylene Blue stock solution.)
4. Place 10ml of the stock solution into a new glass bottle.
5. Dilute stock Methylene Blue solution up to 100ml mark with the 0.1M glycine buffer. This solution is ready to use for vitality tests.
6. Place 500μl of the “ready to use” methylene blue solution into 1.5ml eppendorf tubes for cell & vitality counting.
Note: Discard solution after 6 months.