Term
| Describe how to prepare a smear |
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Definition
1. Thin film of a specimen is aseptically spread on a slide.
2. Film is air dried
3. Heat fixed |
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Term
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Definition
| The procedure that uses dyes to add color to microbes or cellular structures. |
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Term
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Definition
| Chemicals added to microbes. |
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Term
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Definition
| Have positively charged ions that help them adhere to negarively charged bacteria. |
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Term
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Definition
| Have negatively charged ions that are attracted to background. |
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Term
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Definition
-1 dye
-Determines shape and arrangement |
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Term
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Definition
-Places bacteria into categories
EX. Gram Stain and Acid Fast stain |
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Term
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Definition
-Stains specific parts of microbes
Ex.- Endospore stain, negative stain, flagella stain |
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Term
| Know the steps to a Gram Stain |
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Definition
1. Make smear
2. Crystal violet, added for 1 minute
3. Rinse with water
4. Iodine for 1 minute
5. Decolorize with Ethyl alcohol
6. Rinse with water
7. Counterstain with Safranin, added 1 minute
8. Rinse with water
9. Blot |
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Term
| Know the results to a Gram Stain |
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Definition
-Gram postitive bacteria appear purple
Ex-bacillus
-Gram negative bacteria appear red. |
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Term
| Know the Reasoning to the Gram Stain results. |
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Definition
-Gram positive (+) bacteria have thick cell wall and they retain primary stain (crystal violet).
-Gram negative (-) bacteria have thin cell walls that allow decolorization of primary stain and adherence of counterstain (Safranin) |
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Term
| Know the steps to an Endospore Stain |
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Definition
1. make smear
2. add malachite green for 5 minutes over heat
3. cool, rinse with water
4. add safranin for 45 seconds
5. rinse with water
6. Blot dry |
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Term
| Know the results of the Spore stain |
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Definition
-If spores are present they will appear green.
-Spores can be inside or outside the bacteria cell
-Bacteria cell will appear red.
Ex- Bacillus & Clostridium produce spores
Red-Bacteria
Christmas-Spores are present |
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Term
| Know the reasons for the Spore Stain |
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Definition
-Spores are resistant to mild heat but heating for 5 minutes allows spores to open.
-Heat causes spores to absorb primary stain (malachite green) and appear green
-Bacteria cells are decolorized with water and counterstain (safranin) adheres to cells |
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Term
| Know the steps in an Acid Fast Stain |
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Definition
1. Make smear
2. Add carbolfusion for 5 minutes
3. cool, decolorize with
4. rinse with water
5. add methylene blue for 30 seconds
6. Rinse with water
7. Blot dry |
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Term
| Know the steps to an Endospore Stain |
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Definition
1. make smear
2. add malachite green for 5 min. over heat
3. cool, rinse with water
4. Add safranin for 45 seconds
5. Rinse with water
6. Blot dry |
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Term
| Know the results to an endospore stain |
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Definition
-If spores are present they will appear green.
-Spores can be present inside or outside bacteria cell
-Bacteria cell will appear red
EX- Bacillus and clostridium |
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Term
| Know the reasons for the acid fast stain |
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Definition
-some bacteria contain a waxy material called mycolic acid in their cell walls which makes staining difficult
-once hear and carbolfushin are applied acid alcohol doesn't decolorize the bacteria and they appear red
-If the bacteria doesn't contain mycolic acid, carbolfuchin is decolorized with acid alcohol and methylene blue adheres negative. |
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Term
| Know the steps for a negative stain |
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Definition
1. mix bacteria with nigrosin on one end of slide
2. using 2nd slide, spread suspension over entire slide
3. air dry |
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Term
| Know the results of a negative stain |
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Definition
-used to determine prescence of capsule
-capsule are structures that surround the cell wall
-If capsule is present it won't accept the stain and will appear as light spots against a dark background. |
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