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Bio exam 3 Lab #12
Terms in this set (5)
1. Understand what we did and why we did it.
GRAM STAINING AND 3% KOH ALTERNATIVE
2. Be familiar with all of the terms and concepts covered in the lab handout/during the lab.
3. Know how to make a bacterial smear, including all of the steps.
a. Why do you have to let it air dry?
What do you do after it air dries?
What is heat fixing doing exactly?
-less bacteria is better
-not all the way to the slide edges
a. Air dry so Idk says important
b. You heat fix after you air dry
c. Heat fix-
1. Residual water evaporates and cells become more stuck to the slide.
2. The cells are killed.
3. The cells are more ready to take up the stain.
b. What are the steps in Gram Staining? Know the following:
i. What each step is doing specifically.
ii. What order they occur in.
iii. The color the different cell types should be at the end.
1. Crystal Violet Stain- 60 sec
-stick to the peptidoglycan layer of the bacterial cell wall.
2. Iodine- 60sec
-mordant—it combines with the crystal violet to form a larger molecule
3. Acetone/Alcohol Wash-5 sec
-differential step-creates the observable difference:
4.Safranin Stain- 2 min
-stains the cytoplasm of the cell.
4. Be able to answer the questions at the end of the lab handout.
Viscous- gram negative
Non viscous- gram positive
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