Question

Endospore Lab

What is the primary purpose of sporogenesis/ germination

Summarize the spore staining process with these questions

What is used for the primary stain

What cells will become stained spores/vegetative or both?

Whay must heat be added during the primary stain?

What is used as the decolorizing agent?

What does it do?

What cells will become colorless? And why?

What is used for the counterstain?

Which cells will be stained by the counterstain?

At the end of staining process, what is the color of each cell type?

Answer 1

Answer 1) primary purpose:

Sporogenesis - lack of water, lack of food, too much UV light, mutagen. Unfavorable living conditions.

Germination - the presence of water and a specific germination agent. The agent stimulates the formation of hydrolytic enzymes

Answer 3) primary stain:malachite green

Answer 4) both spore become spored:

Endospores: Endospores are bright green.
Vegetative Cells: Vegetative cells are brownish red to pink.

Answer 5)Heat is necessary to force the dye into the endospore wall. Endospores are normally resistant to chemicals and are not easily stained.

Answer 6) decolorizing agent: acid alcohol

97 ml of ethanol, 95% (vol/vol)
3 ml of hydrochloric acid (concentrated)

Answer 8)Endospores: Endospores are bright green.
Vegetative Cells: Vegetative cells are brownish red to pink

Answer 9)Because The addition of a counterstain or secondary stain (safranin) is used to stain the decolorized vegetative cells.

Answer 10)after end of staining:

Endospores: Endospores are bright green.
Vegetative Cells: Vegetative cells are brownish red to pink

Answer 9) safranin is used as counter strain