Spore Stain

Question 1

Step 1 of Spore stain

Answer 1

heat fix organism using Bunsen burner or bacticinerator (like normal)

this is when the bacteria will sporulate d/t the heat stress

Question 2

Step 2 of Spore stain

Answer 2

cover slide with paper towel and then cover paper towel with malachite green; heat using Bunsen burner directly 3-4 seconds 3-4 times; let cool 2 minutes, rinse w/ distilled water

reapply malachite green dye as necessary to avoid drying out

Question 3

Step 3 of Spore stain

Answer 3

cover slide w/ safranin for 1 minute; rinse w/ distilled water

Question 4

Step 4 of Spore stain

Answer 4

blot dry w/ paper towel, observe under 100x lens

Question 5

Method to get paper towel that will “fit” slide.

Answer 5

place slide onto paper towel at the end so that they line up, tear away excess paper towel quickly to leave a perfect slide-sized piece of paper towel

Question 6

What color will spores be?

Answer 6

blueish-green

Question 7

What color will non-sporulating bacteria be?

Answer 7

light pink

(similar to Gram-negative during Gram stain)

Question 8

Why do sporulating bacteria form spores?

Answer 8

to survive harsh environments that “active” cells would die in; specifically to survive high heat and avoid dehydrating

Question 9

What does a vegetative cell look like?

Answer 9

vegetative cellsare the active, growing form of the cell and stain like regular (non sporulating) cells; vegetative cells are what form the endospores