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Microbio Lab > EXAM 1 > Flashcards

EXAM 1 Flashcards

(101 cards)

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1
Q

What is microbiology?

A

The study of organisms that can only be seen well with a microscope.

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2
Q

What are aseptic techniques?

A

Techniques/procedures used to minimize contamination of your sample or work area.

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3
Q

What should you do if something goes wrong in the lab?

A

Tell your professor right away — don’t try to fix it yourself.

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4
Q

Why should long hair be tied back in the lab?

A

To keep it away from flames and chemicals.

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5
Q

What lab safety equipment should you know the location of?

A

Safety shower

Eyewash station

Fire extinguisher

Sink

First aid kit

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6
Q

Why do we use lab safety equipment?

A

To respond quickly to accidents and keep everyone safe.

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7
Q

What does “ubiquity” or “ubiquitous” mean in microbiology?

A

Microorganisms can be found everywhere in the laboratory/environment.

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8
Q

How do you test the ubiquity of microorganisms?

A

1.Take a sterile cotton swab and swab a surface (bench-top, books, computer, hand, floor, etc.)

2.Swab the surface of a Nutrient agar plate

3.Place in incubator at 37°C for 24–48 hours

4.Observe and compare how much and what type of growth occurred

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9
Q

What is nutrient agar used for in this test?

A

It’s a general-purpose growth media for microorganisms.
-Grow a wide variety of bacteria
-Support the growth of non-fastidious organisms (those that don’t need special nutrients)

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10
Q

Why is it important to use aseptic techniques?

A

Because you don’t want to contaminate your sample when working with microorganisms or patient samples.

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11
Q

Why does handwashing matter in microbiology and healthcare?

A

It prevents the spread of germs and helps lower nosocomial infections (infections caught in hospitals).

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12
Q

What is Glo-Germ used for?

A

It’s a lotion with particles that glow under UV light, simulating bacteria for handwashing experiments.

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13
Q

What are the steps in the Glo-Germ handwashing experiment?

A

Apply Glo-Germ on all hand surfaces

View under UV light

Wash hands

View again under UV to see what’s left

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14
Q

What is the proper way to wash hands?

A

Wash for 20 seconds (sing “Happy Birthday” twice).

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15
Q

Why do we use staining in microbiology?

A

To identify bacteria by looking at their shape, arrangement, and size.

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16
Q

What are special stains used for?

A

To see special structures that regular stains can’t reveal.

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17
Q

What are the 4 special staining techniques?

A

1.Flagella staining

2.Capsule staining

3.Endospore staining

4.Acid-fast staining

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18
Q

What are the 4 flagella arrangements?

A

Monotrichous = One flagella

Lophotrichous = Cluster at one end

Amphitrichous = Both ends

Peritrichous = All around the cell

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19
Q

What stains are used for negative staining and why?

A

India ink (bluish) and Nigrosin (reddish) stain the background, not the capsul

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20
Q

Why do we use negative staining for capsules?

A

Capsules don’t stain well, so we stain the background to make them visible.

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21
Q

Steps for negative stain procedure:

A

Drop of Nigrosin + mix bacteria

Air dry (no heat)

View under microscope

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22
Q

What is the purpose of endospore staining?

A

To stain endospores, which are hard to stain because they’re protective and tough.

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23
Q

What are the steps of endospore staining? (we didnt do this one)

A

Heat-fixed smear

Malachite green + steam 12 min

Rinse with water

Safranin 1 min

Rinse, blot dry, view

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24
Q

What is acid-fast staining used for and why?

A

To stain Mycobacterium (like TB) because of their waxy cell wall made of mycolic acid.

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25
Why do we stain microorganisms?
To make them visible and study their shape (morphology), arrangement, and size under a microscope.
26
What are the 3 main bacterial shapes?
Coccus = Spherical Bacillus = Rod Spiral/Curved
27
What are 4 types of cell arrangements?
Single = alone Diplo = in pairs Strepto = in chains Staphylo = grape-like clusters
28
Give examples of shape + arrangement names.
Diplococcus = 2 round cells Streptobacillus = chain of rods Staphylococcus = cluster of round cells
29
What is simple staining?
Using a basic dye (positive charge) to stain the negatively charged surface of a bacterial cell.
30
What stain is commonly used in simple staining?
Methylene Blue, but crystal violet, safranin, and malachite green can also be used.
31
What is the purpose of heat fixing a bacterial smear?
To kill the bacteria and stick it to the slide so it doesn't wash off.
32
What is a chromophore (or chromogen)?
The positively charged part of the stain that sticks to the bacterial cell wall.
33
Steps for a Simple Stain:
1.Make a heat-fixed smear 2.Cover with methylene blue (1 min) 3.Rinse with water 4.Blot dry and view
34
Why is it called Staphylococcus aureus?
Staphylo = grape-like cluster Coccus = round shape
35
How should you carry a microscope?
One hand under the base, one hand holding the arm/neck.
36
What should you do when putting away your microscope?
Set to scanning lens (red band) Lower stage with coarse focus Turn oculars to storage position Wrap cord neatly Clean oil with lens paper + ethanol
37
What type of microscope do we use in lab?
Compound Bright-Field Microscope -Light creates a dark image on a bright background -Uses two lenses (ocular + objective)
38
What is the formula for Total Magnification?
Ocular lens power × Objective lens power Ex: 10× ocular × 40× objective = 400×
39
What does the ocular lens do?
The lens you look through (usually 10x magnification)
40
What are the four objective lenses and their colors?
Scanning = Red (4×) Low Power = Yellow (10×) High Dry = Blue (40×) Oil Immersion = White/Black (100×)
41
What is the Scanning lens used for?
Finding the specimen (not for detailed viewing) Magnification = 4× Total = 40×
42
Why do we use the Low Power lens first?
To focus before switching to higher magnification Magnification = 10× Total = 100×
43
What is the High Dry lens used for and what must you avoid?
Used to zoom in further but still can’t see bacteria clearly Magnification = 40× Total = 400× ⚠️ Never use coarse focus with this lens
44
What lens is used to see individual bacteria clearly?
Oil Immersion lens Magnification = 100× Total = 1000× ⚠️ Do not use coarse focus with this lens either!
45
What is Parcentral Imaging?
If an object is centered at low power, it will stay centered when switching to higher power
46
What happens to your field of view as magnification increases?
It decreases — you see less area but in more detail.
47
What is Parfocal Imaging?
If the object is in focus at low magnification, it will stay almost in focus at higher magnification.
48
When using the high-dry (40x) or oil immersion (100x) lenses, which focus knob should you avoid?
Do NOT use the coarse focus knob — it can break the slide!
49
What is used instead of light in electron microscopes?
Electrons — they allow us to see much smaller details than light microscopes.
50
Are electron microscopes small and cheap?
No, they are very large and expensive.
51
What does a Transmission Electron Microscope (TEM) do?
Produces a 2D image of the inside of the specimen.
52
What does a Scanning Electron Microscope (SEM) do?
Produces a 3D image of the specimen's surface.
53
Which microscope shows interior details of tiny structures?
TEM (Transmission Electron Microscope)
54
Which microscope shows the surface in 3D?
SEM (Scanning Electron Microscope)
55
what is needed to stain the endospore
steam and time
56
Endospores (Made by process called
Sporogenesis
57
endospores can only be make by two types of bacteria
Only made by Bacillus and Clostridium bacteria
58
Autoclaving
the only way to destroy spores High heat (in form of steam) and pressure for 15-20 minutes 121°C & 15 psi (pressure)
59
why is acid fast stainign important
Important because you are trying to figure out which are acid fast positive (contains bacteria causing diseases like TB)
60
Bright-field Microscope
Light passes up through the lens and creates a dark image against a bright background Used to view stained cells
61
Compound microscope
Have 2 sets of lenses: Ocular lens = the one you look through (usually 10x magnification) Objective lens = the lens near the object (usually 3–4 lenses with different powers) Total Magnification (TM) = Ocular power × Objective power Example: 10x (ocular) × 40x (objective) = 400x total magnification
62
compound microscope
Have 2 sets of lenses: Ocular lens = the one you look through (usually 10x magnification) Objective lens = the lens near the object (usually 3–4 lenses with different powers) Total Magnification (TM) = Ocular power × Objective power Example: 10x (ocular) × 40x (objective) = 400x total magnification
63
Binocular lens
Has two oculars (eye piece) -ocular doesnt mean automatically 2
64
what type of cells will produce endospores
vegitative cells
65
what is thing called what we heat and the loop
batic cerator or inoculating loop
66
acid fast staining we did in the lab
Make a heat-fixed smear of Mycobacterium smegmatis on a slide. 2. Cover with CARBOL FUCHSIN, place a paper towel over it, and soak it with more stain. 3. Steam over water bath for 5 minutes, keeping it wet with stain. 4. Remove paper towel, then decolorize with ACID ALCHOL (10–15 seconds). 5. Rinse with water. 6. Stain with METHYLENE BLUE for 60 seconds (counterstain). 7. Rinse with water again. 8. Blot dry, then view under the microscope.
67
what colors does it stain for acid fast negative/positve?
pos - Acid-fast (e.g. TB) - fushia (pink) neg - non acid fast - blue
68
rmb for the microsope portioning:
for microscope portioning: On the first one move the stages slowly until u see something an in the scaling just spin the focus non all around and it it will come
69
What are antiseptics?
Disinfectants that are weakened to be safe for use on living tissues.
70
What’s the purpose of Gram staining?
To identify bacteria and determine if they are Gram-positive or Gram-negative.
71
Who developed the Gram stain, and when?
Hans Christian Gram in the late 1800s.
72
What does Gram staining tell us?
Cell morphology (shape, size, arrangement) and Gram reaction (positive or negative).
73
What are the differences between Gram-positive and Gram-negative bacteria?
G⁺: Thick peptidoglycan, no outer membrane, stains purple/blue G⁻: Thin peptidoglycan, has outer membrane, stains pink
74
What’s the Gram stain procedure?
Heat-fixed smear Crystal violet – 60 sec → rinse Iodine – 60 sec → rinse Alcohol – 10–15 sec → rinse Safranin – 60 sec → rinse Blot dry → microscope
75
What is Gram’s iodine used for?
Acts as a mordant, forming a CV-I complex that locks stain in Gram-positive cells.
76
What is the role of alcohol in Gram staining?
Decolorizes Gram-negative cells by dissolving the outer membrane.
77
What happens when you add safranin in Gram staining?
Stains Gram-negative cells pink/red; Gram-positives stay purple.
78
Antisepsis
Definition: Reduction of microorganisms and viruses, especially pathogens, on living tissue Examples: Iodine, alcohol Comments: Antiseptics are often diluted disinfectants to make them safe for use on tissues.
79
Aseptic
Definition: An environment or procedure free of pathogenic contaminants Examples: Surgical prep, handwashing, flame sterilization Comments: This is a standard practice in labs and healthcare to prevent contamination.
80
-cide / -cidal
Definition: Suffix meaning destruction of a specific type of microbe Examples: Bactericide, fungicide, virucide Comments: Germicides include ethylene oxide, propylene oxide, and aldehydes.
81
Disinfection
Definition: Destruction of most microorganisms and viruses on nonliving tissue Examples: Phenolics, alcohols, aldehydes, soaps Comments: Refers mostly to eliminating pathogens from surfaces.
82
-stasis / -static
Definition: Suffix meaning inhibition (not complete destruction) of a type of microbe Examples: Bacteriostatic, fungistatic, virustatic Comments: Germistatic agents include some chemicals, refrigeration, and freezing.
83
Sterilization
Definition: Destruction of all microorganisms and viruses in or on an object Examples: Preparation of microbiological culture media, canned food Comments: Typically achieved by steam under pressure, incineration, or ethylene oxide gas.
84
Shape (Morphology):
Coccus = round Bacillus = rod-shaped Spirillum = spiral-shaped
85
Arrangement:
Diplo = pairs Strepto= chains Staphylo = clusters
86
autoclave
Temperature: 121°C Pressure: 15 psi (pounds per square inch) Time: 15–40 minutes,
87
gram staining colors
negative: red positive: purple
88
acid fast sating colors
Acid-fast (e.g. TB) - fushia (pink) non acid fast - blue
89
acid fast colors
blue(-)/pink (+)
90
gram colors
red (-)/purple (+)
91
Antisepsis
Definition: Reduction of microorganisms and viruses, especially pathogens, on living tissue Examples: Iodine, alcohol Comments: Antiseptics are often diluted disinfectants to make them safe for use on tissues.
92
Aseptic
Definition: An environment or procedure free of pathogenic contaminants Examples: Surgical prep, handwashing, flame sterilization Comments: This is a standard practice in labs and healthcare to prevent contamination.
93
-cide / -cidal
Definition: Suffix meaning destruction of a specific type of microbe Examples: Bactericide, fungicide, virucide Comments: Germicides include ethylene oxide, propylene oxide, and aldehydes.
94
Disinfection
Definition: Destruction of most microorganisms and viruses on nonliving tissue Examples: Phenolics, alcohols, aldehydes, soaps Comments: Refers mostly to eliminating pathogens from surfaces.
95
-stasis / -static
Definition: Suffix meaning inhibition (not complete destruction) of a type of microbe Examples: Bacteriostatic, fungistatic, virustatic Comments: Germistatic agents include some chemicals, refrigeration, and freezing.
96
Sterilization
Definition: Destruction of all microorganisms and viruses in or on an object Examples: Preparation of microbiological culture media, canned food Comments: Typically achieved by steam under pressure, incineration, or ethylene oxide gas.
97
what contains the all the imoprtant part of cell
endospore
98
Alcohol (Decolorizer): What happens?
In Gram-positive: dye crystals remain trapped in thick peptidoglycan. In Gram-negative: outer membrane is weakened, dye is lost — cell becomes colorless.
99
Gram’s Iodine (Mordant): What does it do?
In Gram-positive: forms a CV-I complex, trapping dye in the thick wall. In Gram-negative: no effect due to outer membrane.
100
Crystal Violet (Primary Stain): What happens?
Both Gram-positive and Gram-negative cells take up the purple dye. It affixes to the cell wall.
101
What does the peptidoglycan layer of a bacterial cell wall look like structurally?
It looks like a crisscrossed network—similar to a chain-link fence—forming one massive molecule that locks the outer structure of the cell into a tight, protective box.

Microbio Lab (4 decks)

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