When antacid staining is used directly on sputum specimens, the thickness of the specimen smear can be increased appropriately to improve the detection rate. When dyeing thick smears, the time of re-staining should be mastered, if the background is too deep, it will affect the microscopic examination. This experiment is from Mudanjiang Medical College, undergraduate 5-year laboratory instruction.
Operation method
Configuration of antacid staining and experiments on staining methods
Principle
When antacid staining is used directly on sputum specimens, the thickness of the specimen smear can be increased appropriately to improve the detection rate. When dyeing thick smears, it is necessary to master the re-staining time, if the background is too dark, it will affect the microscopic examination. Move Alkaline complex red staining: (Cena Ziehl-Neelsen method) For more product details, please visit Aladdin Scientific website.
I. Experimental reagents:
1. CELENA Ziehl-Ziehl-Neelsen solution of carbolic acid:
Alkaline compound red ethanol saturated solution: 10 ml
5% Carbonic acid solution: 90ml
2. Decolorizing agent:
Concentrated hydrochloric acid: 3 ml 95% ethanol: 97 ml
3. Re-staining solution (Lüfler's Melamine solution):
Saturated solution of Melamine in ethanol: 30ml
10% potassium hydroxide: 0.1ml
Distilled water: 100ml
II.Staining method:
l. Smear fixed by flame, add carbolic acid compound red solution, slowly heating until steam appears, never boiling. Dye for 5min, wash with water.
2. Add decolorant, shake the slide from time to time until no red color comes off, wash with water.
3. Add re-staining solution, dye for 0.5-Imin, wash.
4. After drying and microscopic examination, the mycobacteria are red and the background is blue.
Carboniferous Magenta-Melan Sulfate Staining Method
Staining solution:
First solution: carbolic acid magenta solution: take 10ml of basic magenta ethanol saturated solution and add 90ml of 5% aqueous carbolic acid solution and mix.
The second solution: Sulfuric acid Melan solution: Melan 2g, dissolved in 75ml of water, slowly add 25ml of concentrated sulfuric acid and mix to form.
Methods: After the smear is fixed by flame, add the first liquid and dye it with slight heat for 3min, and wash it with water. Then dye with the second solution for about lmin, wash with water. After drying, microscopic examination.
RESULTS: The antacid bacteria were red, and all others were blue.