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90 Cards in this Set
- Front
- Back
What are the two general forms of media?
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1. Liquid (broth)
2. semi-solid (agar) |
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Two forms of general purpose media:
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1. Nutrient broth/agar
2. Tryptic Soy broth/agar |
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The ________ _________ method is the most important isolation tool.
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Streak Plate
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Streaking results in _________ __________ to a single bacterium. The resulting colony will be _______ of one another.
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Limiting dilution (we don't have to know these exact words, but you get the idea)
Clones |
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In lab, we used an agar deep stab to test for ______________.
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motility
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When using oil immersion, only the _____X objective should be used.
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100
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The ______ stain is the most important stain in microbiology.
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Gram
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Gram ____ bacteria appear purple while Gram ____ bacteria appear pink.
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Positive = Purple
Negative = Pink |
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Acid fast stains are primarily used to ID ____________ species.
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Mycobacterial
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The ______ stain is a differential stain based upon ______ _____ composition
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Gram
Cell wall |
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Crystal violet is a _______ dye.
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Basic (as opposed to acidic)
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Gram's Iodine is a fixing agent,also called a ___________.
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Mordant
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Gram's Iodine covalently links crystal violet to a Gram _______ bacteria's cell wall.
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Positive
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_________ alcohol is used to rinse away unbound dye and to __________ Gram negative cells.
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95% ethanol
Decolorize |
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A counter stain, _________ is used to stain Gram ________ bacteria pink.
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Safranin
Negative |
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Mycobacteria have _________ acid in their cell walls, making the ______ _______ staining technique very useful.
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Mycolic Acid
Acid Fast |
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_______ ________ dye binds to mycolic acid and is acid-alcohol resistant.
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Carbol Fuchion
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___________ _________ is the counter stain we used in our acid fast staining procedure.
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brilliant green
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________ Alcohol is the decolorizer in acid fast stain procedure.
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Acid
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List the primary, mordant, decolorizer and counter stain for Gram Staining:
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1. Crystal Violet
2. Gram's Iodine 3. 95% ethanol 4. Safranin |
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List the primary, decolorizer and counter stain for Acid Fast Staining:
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1. Carbol Fuchion
2. Acid Alcohol 3. Brilliant Green |
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Immersion oil constrains __________ (light-scatter)
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Refraction
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What is it and what does it show?
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Good Streak plate showing isolated colonies in the 4th quadrant
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Eukaryote or Prokaryote?
Domain: Division Name: Disease: |
Prokaryote
Domain: Bacteria Division: Cyanobacteria (bluegreen) Name: Anabena (string of pearls) Disease: some produce toxins |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Protista Subkingdom: Protozoa Name: Amoebae proteus Disease: none |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Protista Subkingdom: Protozoa Name: Balantidium coli (note cysts) Disease: Dysentery |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Protista Subkingdom: Protozoa Name: Trypanosoma brucei (gambiense) (Hint with RBCs) Disease: African Sleeping Sickness |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Protista Subkingdom: Protozoa Name: Trichomonas vaginalis Disease: STD |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Protista Subkingdom: Protozoa Name: Plasmodium falciparum (note ring structures in RBCs) Disease: Malaria |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Protista Subkingdom: Algae Name: Spirogyra (multicellular) Disease: none |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Protista Subkingdom: Algae Name: Diatoms (note silica walls) Disease: some produce a neurotoxin - (domoic acid) |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Protista Subkingdom: Algae Name: Ceratium (note central "waist") Disease: Some produce neurotoxins |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Fungi Subkingdom: none Name: Aspergillus (mold) (ball on a stick) Disease: Opportunistic respiratory pathogens |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Fungi Subkingdom: none Name: Penicillium (mold) Flowery fingers) Disease: Opportunistic respiratory pathogens |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Fungi Subkingdom: none Name: Rhizopus (bread mold) (sexual zygospore) Disease:Opportunistic respiratory pathogens |
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Eukaryote or Prokaryote?
Kingdom: Subkingdom: Name: Disease: |
Eukaryote
Kingdom: Fungi Subkingdom: none Name: Rhizopus (bread mold) (asexual sporangiospore) Disease: Opportunistic respiratory pathogens |
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The term that refers to the purposeful
addition of microorganisms into a laboratory nutrient medium is ___________________ |
inoculation
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A pure culture contains only
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one species of microorganism.
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A microbiologist inoculates Staphylococcus aureus into a culture medium. Following incubation, both Staphylococcus aureus and Staphylococcus epidermidis are determined to be growing in this
culture. What is the most likely explanation? |
The culture is contaminated.
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Which of the following characteristics
refers to the microscope's ability to show two separate entities as separate and distinct? |
resolving power
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What feature of a microscope magnifies the
specimen to produce the real image of the specimen? |
objective lens
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If a microbiologist is studying a specimen at a
total magnification of 950X, what is the magnifying power of the objective lens if the ocular lens is 10X? |
95X
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The Gram stain, acid-fast stain, and
endospore stain have the following in common: |
are differential stains (as opposed to simple stains)
Differential stains require two dyes |
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Bacteria that require special growth
factors and complex organic substances are called |
fastidious
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Which two types of media can be used
to determine if a bacteria is motile? |
SIM and TTC
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Why is oil of immersion often used when
viewing specimens under the microscope? |
to increase the resolution
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One colony typically develops from the
growth of several parent bacterial cells. True or False? |
False (one colony - one cell)
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Some microbes are not capable of
growing on artificial media. True or False? |
True (not all critters can be grown in pure cultures or in laboratory conditions.
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The correct microbiological term for the tiny
sample of specimen that is put into a nutrient medium in order to produce a culture is the ____ |
inoculation
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_____ is the term for a culture
made from one isolated colony. |
pure culture
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Newly inoculated cultures must be
_____ at a specific temperature and time to encourage growth. |
incubated
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Magnification is achieved in a compound microscope through the initial magnification of the specimen by the _____ lens. This image is then projected to the _____ lens that will further magnify the specimen to form a virtual image received by the eye.
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objective, ocular
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The _____ of the microscope holds and
allows selection of the objective lenses. |
nosepiece
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_____ dyes have a negative charge on
the chromophore and are repelled by bacterial cells. |
Acidic
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_____ has the same optical qualities as glass
and thus prevents refractive loss of light as it passes from the slide to the objective lens. |
Immersion oil
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Which reagent reacts with crystal violet as the mordant?
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Gram's iodine
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How will E. coli appear if the mordant is not applied?
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Gram-negative
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In lab, Tom was given a mixed culture. His objective is to isolate single colonies. What should be used to accomplish this goal?
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Inoculating loop, Bunsen burner, streak plate
method, 37 degree Celsius incubator |
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What parts of the microscope are used to control the incoming light?
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Iris diaphram
Lamp rheostat |
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How does one determine the magnification of a microscope?
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Multiply ocular power (typically 10X) times the objective power (4X / 10X / 40X / 100X)
10X x 40X = magnification of 400 times |
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What two features does the ocular contain?
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pointer
ocular micrometer |
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What term is used to describe the feature of a microscope in which if one magnification is in focus, all others are in focus when objectives are switched?
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parfocal
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What feature of a microscope focuses and directs the light emitted by the lamp?
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condenser
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What feature of a microscope allows control of the light exiting the condenser?
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Iris diaphram
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What kind of stain is this?
How do you know? |
Acid Fast
Green and pink stains used to stain Mycobaterium |
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What kind of stain is this?
How do you know? |
Mixed Gram
purple and pink stains Gram Pos staphylococci (purple) Gram pos streptobacilli (purple) Gram neg spirillas (pink) |
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What kind of stain is this?
How do you know? |
Negative stain of Micrococcus luteus
looks like an x-ray It is an anionic stain (Nigrosin) that stains the background. It is repelled by the negative surface of the cells allowing you to see the glycocalyx (capsule) |
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What kind of stain is this?
How do you know? Steps of Capsule Stain? |
Capsule Stain ("halos" around cells)
Background is stained with Congo Red (acidic) cells are stained with Manevals' (basic) Capsules remain unstained (neutral charge) |
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What kind of stain is this?
How do you know? |
Flagella Stain
flagella are visible Proteus vulgaris at 1000X showing peritrichous flagella |
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What kind of stain is this?
How do you know? |
Flagella Stain
flagella are visible Spirillium volutans at 1000X showing amphitrichous flagella |
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What kind of stain is this?
How do you know? |
Endospore stain
Bacillus and Clostridium are most common Endospres stain green vegetative cells stain pink |
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What do these tubes show?
What instrument was used to prepare these? What is the medium that used? |
Bacterial motility tests
Inoculation was done with a straight needle stab Motility Test Medium - this one is TTC (tetrazolium salt) (As the bacteria grow, they reduce the TTC, turning it red) |
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What do these tubes show?
What instrument was used to prepare these? What is the medium that used? |
Bacterial motility tests
Inoculation was done with a straight needle stab Motility Test Medium - this one is SIM (SIM does not provide the contrast of TTC, but more "critters" like it.) |
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What do these tubes show?
What is the medium that used? What special indicator does this medium contain? |
Oxygen requirements of microorganisms
FTM (Fluid Thioglycollate Medium) broth Special indicator is resazurin |
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Name what each of the four tubes show.
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1 Anaerobe (no growth at top of tube)
2. Aerobe (growth only at top of tube) 3. Aerotolerant anaerobe (uniform growth throughout) 4. Facultative (better growth at the top, but still growth throughout) |
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What are the steps of an Acid Fast stain?
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1. Primary stain – carbol fuschin
2. decolorizer – acid alcohol 3. counterstain – brilliant green |
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What are the steps of a Gram stain?
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1. Primary stain – Crystal violet
2. Mordant (fixer) – Iodine 3. Decolorizer – 95% ethanol 4. counterstain – safranin |
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What are the steps of an Endospore stain?
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1. Primary stain – Malachite Green
2. Decolorizer – none needed wash with Distilled water 3. counterstain – safranin |
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What are the steps of a Negative stain?
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1. Place a drop of Nigrosin on slide
2. Mix in a tiny sample of bacteria 3. Use a second slide to spread sample across slide 4. Air Dry – NEVER HEAT FIX NEGATIVE STAINS |
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What are the steps of a Smear Prep?
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For organisms grown in broth:
1. spread loopful of culture on slide 2. Heat fix For organisms grown on solid media 1. spread a needleful of culture mixed into a small amount of water 2. Heat fix |
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Capsule Stain
(We did not do in lab. We looked at prepared slides) |
1. Background is stained with (acidic) Congo Red
2. Cells are stained with (basic) Manevals’ Capsules remain unstained and have an unstained “halo” around them. |
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Flagella Stain
(We did not do in lab. We looked at prepared slides) Five Types of Flagella arrangement |
These are difficult to do because the flagella need to be heavily coated with stain.
Know the 5 types: atrichous – no flagella Monotrichous – one flagella Amphitrichous – one at both ends Lophotrichous – flagella tuft at one end Peritrichous – flagella all around |
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Classification of an organism with optimal growth below 15oC
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Psychrophiles
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Classification of an organism with optimal growth below 15oC but as high as 25-30 oC
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Psychrotrophs
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Classification of an organism with optimal growth between 20 - 40oC
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mesophiles
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Classification of an organism with optimal growth above 45oC
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Thermophiles
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What does a spectrophotometer measure?
How did we use it in lab? |
A spectrophotometer measures transmitted (transmittance) or absorbed (absorbance) light at a particular wavelength.
(We used it to measure the turbidity of a broth to determine the amount of bacterial growth. |
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Definition of Diversity
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Many different types of microorganisms may be present in any sample.
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Definition of Ubiquity
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microorganisims are everywhere
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Points to recall when preparing Streak Plate
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1. Label agar side only (name & section #)
2. In tray, store agar side up (lid on bottom) to prevent incoming contaminants or condensation from settling on the agar surface. |