Silver stain for flagella

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Presentation transcript:

Silver stain for flagella Bactria: Escherichia coli Pseudomonas aeruginosa Chapter 15, p 89

objective Stain bacterial flagella which are responsible for bacterial motility

principle Use mordant (stain solution A) to enclose flagella Use sliver stain (stain solution B) to dye the protein of flagella

Solution one (sliver stain A): Stain agents Solution one (sliver stain A): 25 mL of saturated aluminum potassium sulfate (KAl(SO4)2 ) 50 mL of 10% tannic acid 5 mL of 5% ferric chloride (FeCl3) Solution two (sliver stain B): Ammonium hydroxide (AlOH3) 5% sliver nitrate

procedures Clean slide Drop water on the slide Pick up bacterial colony and spread into the drop of water Spread bacterial suspension Air dry (DO NOT use flame) Add solution ONE Stain for 4 min Wash slightly

Add solution TWO Stain for 2 min Place slide on an alcohol burner Do not overheat Heat the slide until steam (about 4 min) Wash Dry Observe under microscope

Pseudomonas aeruginosa

Escherichia coli

Bacillus subtilis

Motility test Bactria: Medium Escherichia coli Staphylococcus aureus TSB + 0.4% agar (4 g/L)(2)

Procedure Stab (about 1 cm) into medium Incubate at 37℃ for 24 hours