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

2. objective
Stain bacterial flagella which are responsible for bacterial motility

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

4. 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

5. 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

6. 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

8. Pseudomonas aeruginosa

9. Escherichia coli

10. Bacillus subtilis

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

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