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65 Cards in this Set
- Front
- Back
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Safety |
-No food or drink -wash hands before and after -personal protective equipment -lab coat, gloves, eye wear, closed toed shoes -disinfect lab bench -dispose of a contaminated material |
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Where do you dispose of test tubes? |
discard rack |
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Where do you dispose of used glass slides? |
disinfectant basin |
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Where do you dispose of used latex gloves or contaminated petri plates? |
biohazard bag (autoclave bag) |
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What does a rheostat do? |
adjust the light intensity |
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What is the condenser knob and condenser? |
concentrates the light beam on the specimen |
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What are the course and fine focus knobs? |
adjust the distance between the stage and objective lens |
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What is the function of the iris diaphragm lever and the base? |
adjust the amount of light coming through the collector lens |
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what does the mechanical stage knob do? |
moves the stage left to right, and back to front stage, stage clip |
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Revolving nose piece |
has 4 objectives lenses -scanning objective: 4X -Low power objective: 10X -High ojective: 40X -Oil immersion: 100X |
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What is the ocular lens? |
the eye pieces -Diopter ring: achieve better focus |
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What are the two things a microscope does? |
-Magnification: ability of a microscope to enlarge an object -Resolution: how well a microscope can show two lines lying close together as 2 distinct lines |
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How do you calculate total magnification? |
ocular (10x) X Objective |
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How to properly clean a microscope? |
-scanning (4X) objective in place -clean slide used -remove excess oil from oil immersion objective -turn down rheostat & turn off light |
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How to use a microscope properly? |
-set up the microscope -centering the condenser -Focusing the microscope on lower ower nad moving up -Focus each eye |
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What is the purpose of oil immersion when using 100X objective? |
minimizes the amount of light refracted or lost |
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How long should you wash your hands for? |
At least 20 seconds of scrubbing |
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Steps to proper hand washing |
-wet hands & lather w/adequate amount of soap -rub palms with fingers interlaces -rub right palm over back of left hand w/fingers interlaced vie versa -hook fingertips together and rub together and rub front and back -clasp right thumb in left palm and scrub vice versa -wash wrists -rinse soap in water -dry w/ paper towel -turn faucet off w/towel |
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What percentage of infection is transmitted by dirty hands? |
80% |
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When is it necessary to wash hands with soap and water instead of using sanitizer? |
When hands are soiled. soap and water are more effective at removing or inactivating microbes. Surfactants in the soap lift soil and microbes from skin. |
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What are three common microbes that can be removed by hand washing? |
-Cryptosporidium -Norovirus Clostridium difficile |
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Ubiquitous |
means microorganisms are found everywhere |
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Pure Culture |
a culture that contains a single organism growing in it |
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Colony |
a group of identical cells derived from a single parent cell |
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Turbidity |
cloudiness in a media that indicated growth |
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Media |
nutrient material suitable for cultivation of microorganisms ( something we use to grow bacteria) |
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incoulum |
a sampling of bacterial culture |
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Aseptic technique |
A way of handling microbes that prevents infection of the handler and others who may be exposed. It is important when handling microbial cultures to prevent contamination. |
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Why to we let the loop cool before obtaining a bacterial sample? |
so you don't kill the microbes & potentially create aerosols. |
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why do we hold test tube with our little finger when taking a sample? |
so you wont place the cap on a surface and then contaminate it |
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why do we obtain a pinpoint amount of inoculum? |
if you take to much then wont have sufficient are on the plate to achieve the dilution needed for seperation of one cell from another |
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What are the step to streak for isolation? |
-cool loop -hold loop like a pencil -rest loop at 45 degree angle -streak in quadrants to ensure entire surface is covers -keep lid on petri plate at all time to prevent contamination -use pinpoint amount of inoculum |
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Why are agar plates labeled at the bottom and inverted when incubated? |
- label the bottom so you can easily read when pulling out of incubation -incubate inverted to prevent condensation from dripping on surface |
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What are some characteristics use to identify a bacterial colony? |
pigment size transparency elevation shape of colony no bacillus or cocci |
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List in order the reagents used in traditional gram stain procedure |
1. crystal violet: primary stain 2. Gram iodine: mordant that combines with crystal violet in cell 3. Acetone-Alcohol: the decolorizer 4. Safranin: counter stain or secondary stain |
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Function of the reagents in gram stain procedure |
1. crystal violet: enters the cell wall of all bacteria 2.combines with the violet to make a bigger cell complex in the wall 3.dissolves the high lipid content on the outer membrane of the Gram Negative cell wall 4. Safranin: stain the Gram negative cell wall since they lost the lipid layer during decoloiztion |
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Why do gram negative cells lose the primary dyr during decolorization? |
Gram negative don't have thick peptidoglycan layer so their outer lipid layer is dissolved and color that the lipid had is gone now |
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Gram stain morphology and results |
Gram + = purple Gram - = pink Morphology -cocci: round, spherical -bacilli: rod shaped -Vibrio: slightly curved rods -spirrillium: stiff spirals |
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Simple Stain vs. Differential Stain |
simple : one dye differential 2 dyes, stain contrasting colors depending on their properties and characteristics |
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Name a genera and the disease is causes |
Mycobacterium Mycobacterium tuberculosis Mycobacterium leprae |
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Identify and acid fast organism under a microscope |
acid fast is pink shapes: cocci or bacillus |
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During acid fast staining how is the primary stain forces into the acid fast bacteria? |
uses heat or chemical to penetrate the cell walls; mycolic acid is what make it hard to penetrate |
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explain why certain organisms are acid fast |
charaacterized by their nearly impermeable cell walls; the glycolipids & mycolic acis in the cell wall of acid fast prevent the alcohol from decolorizing the cell |
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differentiate between decolorizer of acid fast versus gram stain. |
-acid fast: acid alcohol -gram stain- acetone alcohol |
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what are the 3 reagent in acid fast staining? |
1. Carbolfuchsin dye 2. acid alchohol decolorizer 3. methylene blue |
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describe 3 genes located on the pGLO plasmid. |
BLA gene: transcribed continuously codes for beta lactamase Ara-C: transcribed continuously; codes for repressor GFP: regulated by the repressor, will not transcribe unless present of arabinose |
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describe the 3 proteins in pGLO plasmid |
-beta lactamse: breaks apart ampicillin -repressor: regulates the transcription of the GFP gene -Flourescent ptotein: glows under UV light |
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Define competent and how we make E.coli competent |
-competent: bacteria able to take up plasmids -by mixing calcium chloride during their growth phase and by exposing them to extreme heat changes (heat shock) the cell opens up to receive plasmids |
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Explain regulation mechanism of GFP gene |
GPF is regulated by the repressor proein and will not be transcribed without the addition of arabinose. Arabinose will inactivate the repressor protein. |
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Describe different plates used and expected results |
1.Luria Agar- Control E.coli: growth yellowsih color 2. Luria Agar+ Ampicillin - control E.coli: no growth because Am 3. Luria Agar + Ampicillin- pGLOW E.coli: growth BLA amicillin restistan no glow 4. Luria Agar+ Ampicillin+Arabinos- pGLO E.coli: growth and glow because arabinose inactivates the repressor protein; GFP can be transcribes |
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Kirby-Bauer vs Minimal inhibitory Concentration test |
Kirby: to test which antibiotic are likely to affect and pathogen MIC: determine low concentration of antibiotic needed to inhibit growth of an organism. so dosage |
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Beta-lactamase test |
Beta-llactamse destroys penicillin yellow: positive pink: negative |
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media used in atibiotic sensitivity test |
mueler hinton agar plate |
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bateriophage |
a virus that infects and replicates within bacterium |
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Lytic cycle |
one of the 2 cycles of the viral reproduction. results in the destruction of the infected cell and its mebrane |
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Plaque |
and are of clearing in a confluent lawn of bacterial growth, clear are on the place where bacteria have been infected |
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why did we make dilutions of the t-4? |
so we could reach a countable sample |
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calculate the titer of phage in a sample and the dilution of the plate |
#plaques/ dilution * amount of original sample - 43/ (10^5)(.1ml^7) PFY/ml |
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Macroscopic and Microscopic appearance of yeast |
Mac: smooth shiny, raised bumps, smells like bread Mic: cocci or rod shaped |
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Macroscopic and Microscopic appearance of mold |
Mac:fuzzy white smells Mic: areial hyphae, complicated structure |
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Saccharomyces |
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candida albicans |
buds |
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penicillium |
condiospores hyphae |
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rhizopus |
sporangiosprores rounds hyphae |
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Aspergillus |
condiospore balls all around circle hyphae |
