Excersize 10 - Spore Staining

Question 0

Procedure for Dorner method

Answer 0

1) make heavy suspension of bacteria in 5 drops of water
2) add 5 drops of carbolfuchsin
3) heat in beaker of boiling water for 15 min
4) mix several loopfuls in a drop of nigrosin on the slide
5) use another slide to spread the mixture
6) allow to air dry

Question 1

Procedure for Schaeffer-Fulton spore stain method

Answer 1

1) cover smear with paper toweling and saturate it with Malachite green. Steam for 10 minutes
2) after slide has cooled, rinse for 30 sec
3) counter stain with safranin for 1 minute
4) rinse with water
5) blot dry

Question 2

What bacteria is used for its spores?

Answer 2

Bacillus megaterium

Question 3

Schaeffer - Fulton method:
Primary stain
Mordant
Counterstain
Final outcome

Answer 3

Malachite green

Heat

Safranin

Green spore and pink cell

Question 4

Dorner Method:
Primary stain
Mordant
Counter stain 
Final outcome

Answer 4

Carbolfuchsin (intense red)

Heat

Nigrosine (background negative)

Red spore, clear cell and black background

Question 5

Three genera of spore formers

Answer 5

Bacillus

Clostridium

Sporosarcina

Question 6

Diseases of spore formers

Answer 6

Anthrax

Gangrene

Tetanus

Botulism