Term
Microscope Calculation Magnification:
Total Magnification= Magnification by the Objective Lens x Magnification by the Ocular Lens
4x =
10x =
40x =
100x = |
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Definition
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Term
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Definition
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Term
Simple Stains:
Heat fix?
Stain(s)?
Purpose? |
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Definition
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Term
Capsule Stains:
Heat fix?
Stain(s)?
Purpose? |
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Definition
No
Congo Red/Maneval's Stain
Determine the presence of a capsule |
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Term
Gram Stain(s):
Heat fix?
Stain(s)?
Purpose? |
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Definition
Yes
Crystal Violet/Safranin
Gram state |
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Term
Media- TSA & TSB Purpose
Interpretation of Results |
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Definition
General culturing
colonies/turbidity |
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Term
Media- Blood Agar Purpose:
Interpretation of Results: |
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Definition
Detect hemolytic activity
(Beta) hemolysis= complete destruction of RBCs & hemoglobin leading to clearing of media around colonies. (alpha) hemolysis= partial destruction of RBCs producing a greening of the growth. (gamma) hemolysis= No hemolysis characterized by no change in the appearance of media around colonies. |
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Term
Media- Fluid Thioglycollate (FTG) Purpose:
Interpretation of Results: |
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Definition
Determine Aerotolerance
Where (and how well) an organism grows along the gradient tells you its aerotolerance (refer to handout and lab manual). Also contains Resazurin, a color indicator that turns pink when oxidized. |
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Term
[image]
Thioglycollate Medium- Far left, very cloudy |
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Definition
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Term
[image]
Thioglycollate Medium- 2nd one in from the left, Cloudy Spiral down the column |
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Definition
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Term
[image]
Thioglycollate Medium- 3rd one in from the left, faded/opaque streak halfway down the tube. |
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Definition
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Term
[image]
Thioglycollate Medium- 4th one in from the left, thick growth only at the top. |
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Definition
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Term
[image]
Thioglycollate Medium- last tube on the right, only growth is happening in between the upper and lower area of the test tubes. |
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Definition
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Term
Media- MacConkey Agar Purpose:
Interpretation of Results: |
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Definition
Selects for Gram Negative & Indicates Lactose Fermentation
Selects for Gram Negative species (contains Crystal Violet and Bile Salts that inhibit Gram Positive species). Neutral Red Dye- (colorless @ pH>6.8, red @ pH<6.8) provides a color indicator for the fermentation of lactose that is characteristic of Gram Negative bacteria that are members of the Enterobacteriaceae. |
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Term
Phenol Red Broth + Sucrose, Lactose, & Glucose- Purpose:
Interpretation of Results: |
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Definition
Detect carbohydrate fermentation ability by acid/gas formation.
PRB contains phenol red, a color indicator that is yellow below pH6.8, pink to magenta above pH7.4, and red in between. A color change from red to yellow indicates pyruvate or other acids were produced by fermentation. Gas trapped in Durham tube indicates CO2 production by alcohol fermentation. |
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Term
Phenol Red Broth:
Yellow Broth, Bubbles in Tube= |
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Definition
Fermentation with Acid & Gas.
Symbol: A/G |
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Term
Phenol Red Broth:
Yellow Broth, No Bubbles in Tube: |
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Definition
Fermentation with Acid end products, & No Gas produced.
Symbols: A/- |
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Term
Phenol Red Broth:
Red Broth, No Bubbles in Tube= |
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Definition
No Fermentation
Symbols: -/- |
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Term
Phenol Red Broth:
Pink/Orange Broth, No Bubbles in Tube= |
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Definition
Degradation of peptone; alkaline end products
symbol: K |
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Term
| Aseptic Technique Purpose: |
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Definition
| Inoculation technique that prevents contamination of yourself, others, the environment, and the medium being inoculated. |
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Term
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Definition
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Term
| Clean-Slide Prep. Purpose: |
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Definition
| Remove oils and contaminants from a slide. |
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Term
| Bottom up TSA Plate Incubation: |
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Definition
| Prevents moisture that might accumulate on the lid from dripping down on colonies. |
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Term
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Definition
TSB:
1) Using aseptic technique, place a loop-full of culture from TSB in the center/top of the slide marked TSB, swirl it around until it is about the size of a dime.
2) Allow the smear to air dry completely.
3) Using a clothespin as a handle, heat fix the smear by passing the Bottom of the slide through a flame twice slowly but without pausing.
4) Allow the slide to cool.
TSA:
1) Using aseptic technique, place a loop-full of sterile water in the center/top of the slide marked TSA.
2) Use an inoculating needle to gather a small amount of bacteria from a colony on the TSA plate from previous lab.
3) Transfer the bacteria from the needle to the drop of sterile water on the slide by placing the tip of the needle in the water & swirling it around until it is about the size of a dime.
4) Allow the smear to air dry completely.
5) Using a clothespin as a handle, heat fix the smear by passing the Bottom of the slide through a flame twice slowly but without pausing.
6) Allow the slide to cool.
Part 2:
1) Place heat-fixed bacterial smear slides Top up on the slide holder in sink.
2) Use a dropper bottle containing Methylene Blue to cover the smears with stain being Careful Not to Touch the Tip of the Dropper to the Slide!
3) Allow stain to set for 1 minute.
4) Using a clothespin handle, rinse the Bottom of the slides under a gentle stream of water until all excess stain is removed.
5) Allow the stained smears to air dry completely. |
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Term
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Definition
| Methylene Blue is Positively Charged, so it will stick to the negatively charged surface of the cells, so you can see the cells under the microscope. |
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Term
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Definition
1) Using Aseptic Technique, place a Loop-full of Congo Red on the end of the Clean Slide.
2) Using Aseptic Technique, place a Loop-full of Culture from TSB in the drop of Congo Red & Swirl it around to mix and Separate the cells.
5) Take the second slide (the spreader slide) and place it at an angle on the surface of the first slide:
a. Draw the spreader slide into the Congo Red drop.
b. When the drop flows across the width of the spreader slide...
c. Push the spreader slide to the other end painting an even field of dye & bacteria.
6) Allow the smear to air dry (do NOT Heat Fix!)
7) Place the dirty spreader slide in the rack of death.
8) Flood the slide with Maneval's Stain & allow it to stand for 1 minute.
9) Flip the slide over and rinse the bottom.
10) Allow the slide to dry. |
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Term
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Definition
| stains the background around the cells with an acidic stain. |
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Term
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Definition
| the cells themselves are stained. |
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Term
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Definition
| appears as a white "halo" surrounding the cells. |
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Term
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Definition
| Safe Technique used to prevent contamination of the culture, the environment, self, and others. |
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