GRAM STAINING PROCEDURE

Question 1

List down the material needed for gram staining procedure

Answer 1

Microscope slide
Bacterial sample
Bunsen burner (or other heat source)
Crystal violet stain
Gram’s iodine solution
Decolorizer (alcohol, acetone, or a mixture)
Safranin stain
Water
Bibulous paper (or blotting paper)

Question 2

what is the first step during gram staining (smear preparation)

Answer 2

1. Make a thin smear of the bacterial sample on the slide.
2. Heat-fix the smear by carefully passing the slide through a Bunsen burner flame three times

Question 3

What is the second step gram staining

Answer 3

crystal violet staining for 1 minute

Question 4

what is the second solution used in gram staining after crystal violet

Answer 4

Grams iodine for 1 minute

Question 5

what is the significance of grams iodine

Answer 5

acts as a mordant, fixing the crystal violet to the cells

Question 6

what follows after grams iodine

Answer 6

decolorization

Question 7

what solutions are used in decolorization process

Answer 7

ethanol and acetone

Question 8

what follows after decolorization

Answer 8

safranin (counter stain).

Question 9

importance of counterstaining

Answer 9

stains any bacteria that lost crystal violet in the decolorization step.

Question 10

what step is done after every process in the gram staining process

Answer 10

Rinse with water

Question 11

importance of counterstaining (safranin)

Answer 11

Safranin is a red dye that stains bacteria that lost crystal violet during decolorization (Gram-negative).
This step differentiates Gram-negative bacteria from Gram-positive bacteria, which remain purple.