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115 Cards in this Set
- Front
- Back
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Culture: Corynebacterium Diphtheriae
Stain: Methylene Blue Result: Pleomorphic, Rod, Blue, Cationic |
Simple Stain (Basic Dye)
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What charge does Corynebacterium Diphtheriae have when stained with Methylene Blue?
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Cationic (Positive Charge)
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Culture: Corynebacterium Diphtheriae
Stain: Nigrosin Result: Anionic |
Simple Stain (Acid Dye)
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Culture: Bacillus Megaterium
Stain: Nigrosin Result: White cells, dark background |
Negative Stain
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Culture: Staphylococcus Aureus, Mycobacterium Smegmatis, Bacillus Megaterium
Stain: Violet, Iodine, Alchohol and Safranin Result: Purple |
Gram + Stain
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Name the Reagents used in a Gram Stain, in order.
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Crystal Violet, Gram's Iodine, Ethyl Alchohol and Safranin
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What color is a gram + stain?
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Purple.
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What color is a gram - stain?
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Red.
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Name all four bacteria used in gram stains.
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Staphylococcus Aureus, Mycobacterium Smegmatis, Bacillus Megaterium, Pseudomonas Aeruginosa
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In a gram stain, what shape is a Staphylococcus Megaterium?
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Cocci.
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In a gram stain, what shape is a Mycobacterium Smegmatis?
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Rod.
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In a gram stain, what shape is a Bacillus Megaterium?
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Rod.
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In a gram stain, what shape is a Pseudomonas Aeruginosa?
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Rod.
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In a gram stain, what color does Bacillus Megaterium stain?
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Purple.
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In a gram stain, what color does Staphylococcus Aureus stain?
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Purple.
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In a gram stain, what color does Mycobacterium Smegmatis stain?
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Purple.
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In a gram stain, what color does Pseudomonas Aeruginosa stain?
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Red.
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What is the primary stain in a Gram Stain?
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Crystal Violet.
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What is the Mordant in a Gram Stain?
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Gram's Iodine.
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What is the Decolorizer in a Gram Stain?
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Ethyl Alcohol.
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What is the Counterstain stain in a Gram Stain?
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Safranin.
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Name the order of staining techniques in a Gram stain.
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Primary Stain, Mordant, Decolorizer, Counterstain
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In Gram + bacteria, the peptidoglycan layer is thick or thin?
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Thick. The purple crystal violet stain is trapped by the layer of peptidoglycan.
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In Gram - bacteria, the peptidoglycan layer is thick or thin?
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Thin. The pink safranin counterstain is trapped by the peptidoglycan layer.
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Not heat fixing and acidic are characteristics of which stain?
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Negative Stain.
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Heat fixing and the use of Methylene Blue on a smear are characteristics of which stain?
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Simple Stain.
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Which stain is used in a Negative Stain?
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Nigrosin.
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Which stain is used in a Simple Stain?
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Methylene Blue.
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Which bacteria is used in a Spore Stain?
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Bacillus Megaterium.
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The Schaeffer-Fulton Method is used in which type of stain?
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Spore Stain.
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In the Schaeffer-Fulton Method, which stains are used?
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Malachite Green and Safranin.
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In the Schaeffer-Fulton Method, what turns green from the Malachite Green?
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Spore.
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In the Schaeffer-Fulton Method, what turns red from the Safranin?
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Vegetative Cell.
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The Dorner Method is used in which type of stain?
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Spore Stain.
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In the Dorner Method, which stains are used?
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Carbolfuchsin and Nigrosin.
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In the Dorner Method, what turns bacteria red?
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Carbolfuchsin.
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In the Dorner Method, what is colorless and has dark background?
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Nigrosin.
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In the Dorner Method, which bacteria used?
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Bacillus Megaterium.
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What is another name for the Acid Fast stain?
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The Ziehl-Neelson Method.
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What is the bacteria used in the Acid Fast Stain?
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Mycobacterium Smegmatis.
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What is unique about Mycobacterium Smegmatis?
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It's high lipid content.
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Name the Reagents used in an Acid Fast Stain, in order.
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Carbolfuchsin, Heat, Acid Alcohol and Methylene Blue.
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What is the primary stain in an Acid Fast Stain?
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Carbolfuchsin.
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What is the Mordant in an Acid Fast Stain?
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Heat.
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What is the Decolorizer in an Acid Fast Stain?
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Acid Alcohol.
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What is the Counterstain in an Acid Fast Stain?
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Methylene Blue.
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In an Acid Fast Stain, what color is Mycobacterium Smegmatis?
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Red.
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In an Acid Fast Stain, what color is Staphylococcus Aureus?
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Blue.
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In an Acid Fast Stain, is Mycobacterium Smegmatis acid fast +, or -?
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Acid Fast Positive.
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In an Acid Fast Stain, is Staphylococcus Aureus acid fast +, or -?
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Acid Fast Negative.
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What two human diseases are diagnosed using an Acid Fast Stain?
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Mycobacterium Tuberculosis and Mycrobacterium Leprae.
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What does E.Coli look like on an MSA plate and why?
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Clear (no growth); not salt tolerant
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What does S. Epidermidis look like on an MSA plate and why?
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White, due to growth
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What does Staphylococcus Aurerus look like on an MSA plate and why?
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Yellow, due to fermentation of Mannitol
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What is the optimum temperature for Serratia Marcescens?
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25 degrees C.
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What is the pigment produced in Serratia Marcescens?
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Prodigiosin.
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In MSA plate, why is bacteria differential selective?
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Because of High Salt Concentration.
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What is the organism that grows at high salt concentration?
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Halobacterium Salinarium (Obligate Halophile).
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Is Halobacterium an obligate halophile or facultative halophile?
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Obligate Halophile.
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What is the organism that grows at widest range of salt concentration?
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Staphylococcus Aureus.
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What is the organism that grows at smallest range of salt concentration?
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E. Coli.
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Most Effective Wavelength of UV light to kill bacteria?
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260 nm, DNA maximally absorbs UV light forming pyrimidine dimers
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What does DNA form when absorbing UV light?
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Pyrimidine dimers.
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Why are lids removed before UV exposure?
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UV rays cannot penetrate solids very well.
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What is the control in the UV light experiment?
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Index card.
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Why are UV Ray exposure times different for different organisms?
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Some like Bacillus Megaterium (spore forms) are more resistant to UV
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What is the Kirby-Bauer Method?
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Antimicrobic Sensitivity Testing.
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Name cultures used in Kirby-Bauer Method.
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S. Aureus, E. Coli, P. Vulgaris, P. Aeruginosa
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What is the medium used in Kirby-Bauer Method?
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Mueller-Hinton II Agar (MHA)
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What is the optimum pH in Kirby-Bauer Method?
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7.2 – 7.4
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What is the optimum Thickness in Kirby-Bauer Method?
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Uniform thickness of 4mm w/ 5% Sheep’s blood
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How are zones of inhibition measured?
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Measured in millimeters b/w edge of disc and growth in 3 categories.
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What are the 3 zones of inhibition?
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1) Resistant less than or equal to 14 mm
2) Intermediate 15 – 16 mm 3) Sensitive greater than or equal to 17mm |
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What is the range for RESISTANT zone of inhibition for discs?
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Less than or equal to 14 mm
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What is the range for INTERMEDIATE zone of inhibition for discs?
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15 – 16 mm
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What is the range for SENSITIVE zone of inhibition for discs?
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Greater than or equal to 17mm
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What method evaluates antiseptics?
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The Filter Paper Disk Method.
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What cultures are used for the Filter Paper Disk Method?
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S. Aureus and P. Aeruginosa
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What is the result of S. Aureus in the Filter Paper Disk Method?
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Gram +, has 2 layers, easy to use disinfectant
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What is the result of P. Aeruginosa in the Filter Paper Disk Method?
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Gram -, has 3 layers, harder to use disinfectant
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In a Brightfield microscope, what does the condenser contain?
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Diaphragm
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In a Brightfield microscope, what functions the light source?
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Voltage Control; it varies light intensity
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In a Brightfield microscope, what functions the Ocular Adjusment?
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Diopter Adjustment Ring (located around the oculars)
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What condenser setting is used for Brightfield?
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The highest position to allow maximum amount of light to enter objective lens.
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Name 4 ways to improve resolution in a Brightfield.
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1) Blue filter placed over light source to decrease wavelength (provides maximum resolution)
2) Condenser should be placed at highest position 3) Not close diaphragm down too much (Close diaphragm improves contrast) 4) Immersion oil at 100X |
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How do you calculate Total Magnification?
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Multiplying ocular lens times objective lens.
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What is the best Focusing Procedure?
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Start with the low-power objective and progress to the higher magnifications
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What is the Oil Immersion Theory?
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Oil immersion reduces light diffraction (scattering) and maximizes the numerical aperture to improve the resolution.
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When do you use Darkfield over Brightfield?
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To see transparent living organisms.
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What are the Condenser Settings used on a Darkfield Microscope?
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a) Star diaphragm
b) cardioid condenser c) paraloid |
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What human disease is often diagnosed with Darfield?
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Syphillis.
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When do you use Phase Contrast Microscopy?
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To see transparent protoplasmic structures and enhance the contrast b/w a cell and its surroundings, without staining. Microscope of choice to view motility.
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What organisms are typically studied using Phase Contrast Microscopy?
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Motile organisms.
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What is the Cell shape for Bacilli?
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Rod
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What is the Cell shape for Spherical?
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Cocci
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What is the Cell shape for
Spiral? |
Curved Rods.
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What are the three Cell Shapes?
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Rod
Cocci Spiral |
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How do you label Petri Plates?
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Label bottom (agar) with initials, assignment number and date
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What property of agar is ideal for solidifying agent?
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Polysaccharide
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What temperature does it take for Agar to melt?
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Melts at 100 ̊C.
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What temperature does it take for Agar to solidify?
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Solidifies at 45 ̊C – 50 ̊C.
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What is the Agar pour technique?
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1) Liquefy agar by boiling in beaker for 5 min
2) Cool down to 50 ̊C for 5 min 3) Remove cap from tube and flame 4) Pour into Petri plate |
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What are the 3 Streak Plate Methods?
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1) Quadrant Streak
2) Radiant Streak (Looks like a ray) 3) Continuous streak (Squiggle looking) |
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Why are your agar plates incubated upside down?
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To prevent condensation and contamination.
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In Smear Prep, what is the Advantage of heat fixing?
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Adhere to organism to slide.
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In Smear Prep, what is the Disdvantage of heat fixing?
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Shrinkage; cell death (motility)
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What does the FTM Fluid Thioglycollate Medium (Tube) contain?
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Glucose, Cystine and Sodium Thioglycollate
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What reduces oxidation/reduction requirements?
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Glucose, Cystine and Sodium Thioglycollate
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What is the INDICATOR in th FTM Fluid Thioglycollate Medium (Tube)?
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Resazurin, indicator of oxygen that becomes pink
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What color is Resazurin because of oxygen?
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Pink.
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What is a Thermal Death Point?
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The temperature in which an organism is killed in 10 min.
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What is a Thermal Death of Time?
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Time required to kill at a given temperature.
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What Subculture Technique do you use for Serial Dilution?
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Pour plate method.
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What Subculture Technique do you use for Streak Plate?
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Primary (mixed culture) to secondary; isolate organism; Test tube to Plate (Get colony from plate and put on another plate (2ndary)
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Do you Heat-Fix in a Smear Prep?
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Heat-Fixed is required.
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