Describe what happens at each step in the gram stain

Asked by admin @ 01/12/2021 in Biology viewed by 274 People

List the steps of the gram stain technique and describe the purpose of each. be sure to include a description of the color of the bacteria during each step.

Answered by admin @ 01/12/2021

The procure of gram staining is as follows:

1. Prepare the smear on a microscope slide and heat-fix it

2. Flood the smear with crystal violet on a staining tray

3. Gently and lightly rinse the smear with sterile water from a wash bottle

4. Flood the smear with iodine for a minute

5. Rinse the smear like in step 3

6. Pour 95% ethyl alcohol or acetone to decolorize for 10 seconds

7. Rinse like in step 3

8. Flood the smear with safranin (a counter-stain) for about 1 minute

9. Rinse the smear like in step 3

10. Leave the smear to air dry and observe under a microscope

Heat-fixing adheres the smear to the slide and hence makes it less amenable to being washed away during gram-staining procedure. Gram staining using crystal violet allows the peptidoglycan layer of the Gram + bacteria to pick up the dye and retain it even after decolorizing. Gram- bacteria lose the crystal violet color after decolorizing with alcohol since they lack the peptidoglycan layer. Safranin is a counter stain since it is picked by the Gram – bacteria that have lost the crystal violet dye. Therefore under a microscope and after the staining procedure, Gram+ bacteria appear violet/purple in colr while Gram – appear pink.

Describe what happens at each step in the gram stain

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