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33 Cards in this Set

  • Front
  • Back
A device that replaces eyepiece of microscope used to measure things.
ocular micrometer
Write the entire process of cleaning and putting away a microscope.
Answer

Write the entire process of cleaning and putting away a microscope.
What are ways to increase resolution when using microscope?
Blue Filter - short wavelength of blue light provides maximum resolution

The Condensor - kept at highest position to allow maximum amount of light into objective.

The diaphragm - should not be stopped down to much.

Immersion oil - used for 100x objective.
What 2 stains bind bile salts?
Cholestipol, Cholestyramine
The 3 lens systems?
Oculars

Objectives

Condenser
What is the formula for ocular micrometer calibration?
Each ocular division =

stage micrometer
_______________
ocular micrometer

then multiplied by 0.01 mm
then convert to um (micrometers)
Briefly describe negative staining
Mixing microorganisms in a small amount of nigrosine (dark purple) and spreading the mixture over the surface of the slide.

Leaves organisms transparent and visible in a darkened field.
Briefly describe the 2 techniques for negative staining.
1) Organisms dispersed onto small drop of nigrosine. Spreader is used to spread across slide. Air dried.

2) Loopful of nigrosine is placed inside of target circle. Organisms added to nigrosine and then spread around to fill target circle. Air Dried.
What where the two control organisms used for Experiment 9: Negative Staining?
Staphylococcus aureus and Bacillus megaterium.
What stain is used for Negative Staining?
Nigrosine or India Ink. (Dark Purple)
Describe briefly what Negative Stain looks like under microscope?
Dark purple background with clear organisms.
Briefly describe smear preperation for both Liquid Media and Solid Media.
Initial slide and draw target circle. Shake culture. Transfer two loop fulls of Liquid media over target circle. Air dry. Heat fix.

If solid media, two loopfuls of water in target circle, then very small amount of organism with inoculating needle. Air dry. Heat fix.
Briefly describe simple staining.
Prepare a slide (normal smear preparation).
Stain with methylene blue for one minute.
Wash off with water.
Blot dry in bibulous paper.
What stain is used in simple staining for 1 minute before washing off?
Methylene blue
This is used to dry slides by blotting.
Bibulous paper
What two organisms were used in simple staining?
Bacillus magaterium
Staphyloccocus aureas
Explain acid fast bacteria
Acid fast bacteria are those such as Mycobacterium tuberculosis and Mycobacterium leprae. They have outer membranes covered with waxy mycolic acid.
Briefly describe capsule staining.
Two loopfuls of organism are mixed in a drop of Nigrosine.
Spread over a slide and air dried.
Heat fixed.
Stained with crystal violet for one minute then washed off.
Blotted with bibulous paper.
What does capsule staining look like under a microscope?
Dark purple background. Clear capsule with purple organism inside. Essentially a purple organism with a clear halo around it.
What was the positive control organism used for capsule staining?
Klebsiella pneumoniae
(coccus)
Briefly describe gram staining process.
Crystal Violet - 20 seonds
Wash
Iodine (mordant) - 1 minute
Decolorize with alcohol (takes color away from gram negative)- 10-20 sec
Wash
Safranin - 20 seconds
Wash
Blot Dry
What does gram staining look like under microscope?
Gram positive organisms (purple)
Gram negative organisms (pink)
What where the control organisms for gram staining?
Eserichia Coli (E. Coli) - Gram negative

Mycobacterium smegmetis - Gram positive

Staphylococcus Aureus - Gram positive
Know these 3 Gram Positive Organisms
Bacillus megatarius
Microbacterus mcmadus
Staphlococcus aureus
Briefly describe Endospore (spore) staining. Schaeffer-Fulton method.
Bring water to boil in a beaker over hot plate.
Cover smear with small piece of paper towel and saturate with malachite green. Steam for 5 minutes.

Remove paper towling and rinse with water for 30 sec.

Counter-stain with safranin for 20 seconds.

Rinse with water

Blot dry with bibulous
Briefly describe Endospore (spore) staining. Dorner Method.
Disperse several loopfuls of bacteria in 5 drops of sterile water (all in a test tube)

Add 5 drops of carbolfuchsin

Heat test tube in a beaker of boiling water for 10 minutes.

Mix several loopfuls of bacteria in a drop of nigrosine on a slide.

Spread nigrosine-bacteria mixture using a slide.

Allow smear to air dry.
What will Endospore staining of Dorner Method look like under microscope?
Red Endospore
Clear Cell
What will endospore staining using the Schaeffer-Fulton method look like under a microscope?
Green Endospore
Pink Cell
Briefly describe the steps of Acid Fast staining.
Bring beaker of water to boil on hot plate.
Place prepared smear on beaker and cover with carbolfuchsin.
Boil for 5 minutes.
Decolorize with acid-alcohol for 15-20 seconds.
Rinse with water
Counterstain with methylene blue for 30 seconds
Rinse with water
Blot dry with bibulous paper
What does Acid Fast Staining look like under a microscope?
Acid fast organisms will be RED (carbolfuchsin stains waxy lipid coating)

Non-Acid fast organisms will be BLUE (Methylene blue)
What is another name for Acid Fast Staining method?
Ziehl Neelsen Method
What are the controls for Acid Fast staining?
Positive - Mycobacteria Mcmanus
Negative - Staphlococcus aureus
Describe motility determination experiment.
Wire with organism is brought into agar test tube without touching walls.
Wire penetratres medium to two thirds of it's depth
Wire is withdrawn and tube is flamed and plugged.