1. Practical seminar 4 Acid fast staining - Mycobacterium tuberculosis
Corynebacteriuôm diphtheriae – Albert´s staining of metachromatic granules

2. Acid fast staining
Bacterial cell resistant to colorisation with some staining (Gram)
Their cell wall is relatively not permeable, containig lipids, fatty acids...(ex. Mycobacterium tbc, Corynebacteria...)
If stained (with strong basic stains (basic fuchsin in 5% fenole – they resist to decolorisation with strong acids (20% H2SO4)

3. Ziehl Neelsen staining
Heat fixed smear
Apply carbol fuchsin (basic fuchsin + phenol)
Warm untill evaporation
Rinse with water
Apply 3% HCl (or 95% alcohol
Counterstain with malachit green

4. Mycobacterium tbc – farbenie podľa Ziehl Neelsena

6. Metachromatic granules staining
Volutin storage granules in cytoplasma
Stained with basic stain
Toluidin blue, methylen blue
They are acidoresistent
Differentiation of C. difteriae from saprophytic corynebacteria
Albert´s staining

8. Metachromatic staining - principals
Stain aplied on fixed cell or tissue is of different color as is the result color
Ernst Babes inclusions – metachromatic granuls – volutine granuls – phosphate molecules - Corynebacterium diphteriae
Volutin – nucleoprotein complex – in cytoplasm of some bacteria, molds and parasits in the forme of storage granules

9. Albert´s staining Fix smear Apply Albert´s solution Add Lugol solution
Bacteria are green or blue, (malachit green or methylen blue), granuls are red