Primary stain crystal violet
01.What color is each cell after Crystal Violet is added?
Gram positive bacteria retains crystal violet
Gram negative bacteria is red or pink coloring.
Mordant:Iodine added to cell. Crystal Violet forms ionic bond with Iodine to form CV-I Complex
02. What color is each cell after added iodine?
cells appear colorless.
Decolorized is added to cell
03.What color is each cell after decolorized is added
the gram-positive cell remains purple in color
gram negative cell loses the purple color and its only revealed when counterstain.
Counterstain, Safrain is added to cell
04.What color is each cell after safranin is added
Gram-postiive: does not disrupt color purple coloration
Gram-negative: cell stain red
Step1: Primary stain Carbol fuchsin is added to each cell, this stain has an additional chemical, phenol, that helps the stain penetrate
5A. What and where is the mordant
-A mordant is a chemical or physical agent that enhances the primary stain
What is a mordant: Primary stain that binds with crystal violet
-
5b:What color is each cell?
Gram Positive: dark crystal violet instead of the safranin cells appear purple
Gram Negative: after ethanol it appears red
5c:Identify the fast acid + and fast acid -
Acid fast positive cells stain bright pink/magenta and are members of genus Mycobacterium
Acid fast negative cells stain blue and are not members of the genus Mycobacterium
Step2: Decolorizer is added to the cell
6A:What is decolorizer made of?
Ethanol
6B:What color is each cell?
Pink
Step3: Counterstain, methylene blue is added to the cell
7A:What color is each cell?
Pink
7B:Identify The acid fast cell
Used to identify acid fast organisms such as members of the genus family mycobacterium
7C:What else do you know about the Acid fast cell?
Acid fast smear will have a mix of both pink and blue cells
7D.What disease may be identified
Mycobacterium
Slide placed over hotplate for 5 mins. This heat is just strong enough to loosen the Peptidoglycan and protein layers surrounding the spore to allow the stain into the spore.
8A:What is the mordant
Steam
8B:What color is the cell?
Red
8C:What does the small circle In the cell represent?
Green endospores
8D:What color is that small circle?
Green
Spore Staining
9A:What is decolorized made of
Water
9B:What is the color of the cell after decolorized is added?
Malachite Green
9C:What color is of the small circle after decolorized is added?
red
10A:What is the color of the cell after Safranin is added?
Cell stain red
10B:What color is the small circle after safranin is added
green
10C:What do you know about who these cells might be?
Bacillius or clostridium
10D:What type of environment were these cells in? How do you know ?
-These bacteria often live in soil, so it could be. a natural stress such as a drought that initiates their conversion into an endospore
-Artificial stress, such as treatment with desinfectants, would have a similar effect.
10E: What disease may be seen with this
Bacillius, clostridium
11A:What are the clear circles?
Capsule
11B.What are the tiny dark circles?
Bacterium
11C:What do you know about the cells with the clear circles compared to the cells with no clear circle?
-Capsule will resist stain and will make the cells that have capsule visible as white areas against the dark background
-cells without a capsule will take in the dark stain and will match the background.