Diana's Micro Lab Test 1

Front 1

What are the two general forms of media?

Back 1

1. Liquid (broth)
2. semi-solid (agar)

Front 2

Two forms of general purpose media:

Back 2

1. Nutrient broth/agar
2. Tryptic Soy broth/agar

Front 3

The ________ _________ method is the most important isolation tool.

Back 3

Streak Plate

Front 4

Streaking results in _________ __________ to a single bacterium. The resulting colony will be _______ of one another.

Back 4

Limiting dilution (we don't have to know these exact words, but you get the idea)
Clones

Front 5

In lab, we used an agar deep stab to test for ______________.

Back 5

motility

Front 6

When using oil immersion, only the _____X objective should be used.

Back 6

100

Front 7

The ______ stain is the most important stain in microbiology.

Back 7

Gram

Front 8

Gram ____ bacteria appear purple while Gram ____ bacteria appear pink.

Back 8

Positive = Purple
Negative = Pink

Front 9

Acid fast stains are primarily used to ID ____________ species.

Back 9

Mycobacterial

Front 10

The ______ stain is a differential stain based upon ______ _____ composition

Back 10

Gram
Cell wall

Front 11

Crystal violet is a _______ dye.

Back 11

Basic (as opposed to acidic)

Front 12

Gram's Iodine is a fixing agent,also called a ___________.

Back 12

Mordant

Front 13

Gram's Iodine covalently links crystal violet to a Gram _______ bacteria's cell wall.

Back 13

Positive

Front 14

_________ alcohol is used to rinse away unbound dye and to __________ Gram negative cells.

Back 14

95% ethanol
Decolorize

Front 15

A counter stain, _________ is used to stain Gram ________ bacteria pink.

Back 15

Safranin
Negative

Front 16

Mycobacteria have _________ acid in their cell walls, making the ______ _______ staining technique very useful.

Back 16

Mycolic Acid
Acid Fast

Front 17

_______ ________ dye binds to mycolic acid and is acid-alcohol resistant.

Back 17

Carbol Fuchion

Front 18

___________ _________ is the counter stain we used in our acid fast staining procedure.

Back 18

brilliant green

Front 19

________ Alcohol is the decolorizer in acid fast stain procedure.

Back 19

Acid

Front 20

List the primary, mordant, decolorizer and counter stain for Gram Staining:

Back 20

1. Crystal Violet
2. Gram's Iodine
3. 95% ethanol
4. Safranin

Front 21

List the primary, decolorizer and counter stain for Acid Fast Staining:

Back 21

1. Carbol Fuchion
2. Acid Alcohol
3. Brilliant Green

Front 22

Immersion oil constrains __________ (light-scatter)

Back 22

Refraction

Front 23

What is it and what does it show?

Back 23

Good Streak plate showing isolated colonies in the 4th quadrant

Front 24

Eukaryote or Prokaryote?
Domain:
Division
Name:
Disease:

Back 24

Prokaryote
Domain: Bacteria
Division: Cyanobacteria (bluegreen)
Name: Anabena (string of pearls)
Disease: some produce toxins

Front 25

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 25

Eukaryote
Kingdom: Protista
Subkingdom: Protozoa
Name: Amoebae proteus
Disease: none

Front 26

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 26

Eukaryote
Kingdom: Protista
Subkingdom: Protozoa
Name: Balantidium coli (note cysts)
Disease: Dysentery

Front 27

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 27

Eukaryote
Kingdom: Protista
Subkingdom: Protozoa
Name: Trypanosoma brucei (gambiense) (Hint with RBCs)
Disease: African Sleeping Sickness

Front 28

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 28

Eukaryote
Kingdom: Protista
Subkingdom: Protozoa
Name: Trichomonas vaginalis
Disease: STD

Front 29

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 29

Eukaryote
Kingdom: Protista
Subkingdom: Protozoa
Name: Plasmodium falciparum (note ring structures in RBCs)
Disease: Malaria

Front 30

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 30

Eukaryote
Kingdom: Protista
Subkingdom: Algae
Name: Spirogyra (multicellular)
Disease: none

Front 31

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 31

Eukaryote
Kingdom: Protista
Subkingdom: Algae
Name: Diatoms (note silica walls)
Disease: some produce a neurotoxin - (domoic acid)

Front 32

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 32

Eukaryote
Kingdom: Protista
Subkingdom: Algae
Name: Ceratium (note central "waist")
Disease: Some produce neurotoxins

Front 33

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 33

Eukaryote
Kingdom: Fungi
Subkingdom: none
Name: Aspergillus (mold) (ball on a stick)
Disease: Opportunistic respiratory pathogens

Front 34

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 34

Eukaryote
Kingdom: Fungi
Subkingdom: none
Name: Penicillium (mold) Flowery fingers)
Disease: Opportunistic respiratory pathogens

Front 35

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 35

Eukaryote
Kingdom: Fungi
Subkingdom: none
Name: Rhizopus (bread mold) (sexual zygospore)
Disease:Opportunistic respiratory pathogens

Front 36

Eukaryote or Prokaryote?
Kingdom:
Subkingdom:
Name:
Disease:

Back 36

Eukaryote
Kingdom: Fungi
Subkingdom: none
Name: Rhizopus (bread mold) (asexual sporangiospore)
Disease: Opportunistic respiratory pathogens

Front 37

The term that refers to the purposeful
addition of microorganisms into a laboratory
nutrient medium is ___________________

Back 37

inoculation

Front 38

A pure culture contains only

Back 38

one species of microorganism.

Front 39

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?

Back 39

The culture is contaminated.

Front 40

Which of the following characteristics
refers to the microscope's ability to show two
separate entities as separate and distinct?

Back 40

resolving power

Front 41

What feature of a microscope magnifies the
specimen to produce the real image of the
specimen?

Back 41

objective lens

Front 42

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?

Back 42

95X

Front 43

The Gram stain, acid-fast stain, and
endospore stain have the following in
common:

Back 43

are differential stains (as opposed to simple stains)
Differential stains require two dyes

Front 44

Bacteria that require special growth
factors and complex organic substances are
called

Back 44

fastidious

Front 45

Which two types of media can be used
to determine if a bacteria is motile?

Back 45

SIM and TTC

Front 46

Why is oil of immersion often used when
viewing specimens under the microscope?

Back 46

to increase the resolution

Front 47

One colony typically develops from the
growth of several parent bacterial cells.
True or False?

Back 47

False (one colony - one cell)

Front 48

Some microbes are not capable of
growing on artificial media. True or False?

Back 48

True (not all critters can be grown in pure cultures or in laboratory conditions.

Front 49

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 ____

Back 49

inoculation

Front 50

_____ is the term for a culture
made from one isolated colony.

Back 50

pure culture

Front 51

Newly inoculated cultures must be
_____ at a specific temperature and time to
encourage growth.

Back 51

incubated

Front 52

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.

Back 52

objective, ocular

Front 53

The _____ of the microscope holds and
allows selection of the objective lenses.

Back 53

nosepiece

Front 54

_____ dyes have a negative charge on
the chromophore and are repelled by
bacterial cells.

Back 54

Acidic

Front 55

_____ has the same optical qualities as glass
and thus prevents refractive loss of light as it
passes from the slide to the objective lens.

Back 55

Immersion oil

Front 56

Which reagent reacts with crystal violet as the mordant?

Back 56

Gram's iodine

Front 57

How will E. coli appear if the mordant is not applied?

Back 57

Gram-negative

Front 58

In lab, Tom was given a mixed culture. His objective is to isolate single colonies. What should be used to accomplish this goal?

Back 58

Inoculating loop, Bunsen burner, streak plate
method, 37 degree Celsius incubator

Front 59

What parts of the microscope are used to control the incoming light?

Back 59

Iris diaphram
Lamp rheostat

Front 60

How does one determine the magnification of a microscope?

Back 60

Multiply ocular power (typically 10X) times the objective power (4X / 10X / 40X / 100X)

10X x 40X = magnification of 400 times

Front 61

What two features does the ocular contain?

Back 61

pointer

ocular micrometer

Front 62

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?

Back 62

parfocal

Front 63

What feature of a microscope focuses and directs the light emitted by the lamp?

Back 63

condenser

Front 64

What feature of a microscope allows control of the light exiting the condenser?

Back 64

Iris diaphram

Front 65

What kind of stain is this?
How do you know?

Back 65

Acid Fast
Green and pink stains
used to stain Mycobaterium

Front 66

What kind of stain is this?
How do you know?

Back 66

Mixed Gram
purple and pink stains
Gram Pos staphylococci (purple)
Gram pos streptobacilli (purple)
Gram neg spirillas (pink)

Front 67

What kind of stain is this?
How do you know?

Back 67

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)

Front 68

What kind of stain is this?
How do you know?
Steps of Capsule Stain?

Back 68

Capsule Stain ("halos" around cells)
Background is stained with Congo Red (acidic)
cells are stained with Manevals' (basic)
Capsules remain unstained (neutral charge)

Front 69

What kind of stain is this?
How do you know?

Back 69

Flagella Stain
flagella are visible
Proteus vulgaris at 1000X showing peritrichous flagella

Front 70

What kind of stain is this?
How do you know?

Back 70

Flagella Stain
flagella are visible
Spirillium volutans at 1000X showing amphitrichous flagella

Front 71

What kind of stain is this?
How do you know?

Back 71

Endospore stain
Bacillus and Clostridium are most common
Endospres stain green
vegetative cells stain pink

Front 72

What do these tubes show?
What instrument was used to prepare these?
What is the medium that used?

Back 72

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)

Front 73

What do these tubes show?
What instrument was used to prepare these?
What is the medium that used?

Back 73

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.)

Front 74

What do these tubes show?
What is the medium that used?
What special indicator does this medium contain?

Back 74

Oxygen requirements of microorganisms
FTM (Fluid Thioglycollate Medium) broth
Special indicator is resazurin

Front 75

Name what each of the four tubes show.

Back 75

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)

Front 76

What are the steps of an Acid Fast stain?

Back 76

1. Primary stain – carbol fuschin
2. decolorizer – acid alcohol
3. counterstain – brilliant green

Front 77

What are the steps of a Gram stain?

Back 77

1. Primary stain – Crystal violet
2. Mordant (fixer) – Iodine
3. Decolorizer – 95% ethanol
4. counterstain – safranin

Front 78

What are the steps of an Endospore stain?

Back 78

1. Primary stain – Malachite Green
2. Decolorizer – none needed wash with Distilled water
3. counterstain – safranin

Front 79

What are the steps of a Negative stain?

Back 79

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

Front 80

What are the steps of a Smear Prep?

Back 80

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

Front 81

Capsule Stain
(We did not do in lab. We looked at prepared slides)

Back 81

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.

Front 82

Flagella Stain
(We did not do in lab. We looked at prepared slides)

Five Types of Flagella arrangement

Back 82

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

Front 83

Classification of an organism with optimal growth below 15oC

Back 83

Psychrophiles

Front 84

Classification of an organism with optimal growth below 15oC but as high as 25-30 oC

Back 84

Psychrotrophs

Front 85

Classification of an organism with optimal growth between 20 - 40oC

Back 85

mesophiles

Front 86

Classification of an organism with optimal growth above 45oC

Back 86

Thermophiles

Front 87

What does a spectrophotometer measure?

How did we use it in lab?

Back 87

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.

Front 88

Definition of Diversity

Back 88

Many different types of microorganisms may be present in any sample.

Front 89

Definition of Ubiquity

Back 89

microorganisims are everywhere

Front 90

Points to recall when preparing Streak Plate

Back 90

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.