Topic 6 Flashcards

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1
Q

What Antonie Van Leeuwnhoek known for?

A

“Animalacules”

- observed single cells in pond water

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2
Q

Dissecting Scope Features (6)

A
  • light reflected off specimen
  • larger depth of field
  • room to manipulate specimen
  • see 3-dimensional image
  • image not inverted
  • lower magnification than compound scope
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3
Q

Features of the Compound Light Microscope

A
  • Ocular and Objective Lenses
  • Light passes through the specimen
  • The image is inverted
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4
Q

What is the magnification of the compound light microscope?

A

up to 1000x

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5
Q

What does oil immersion do in the compound light microscope?

A

Oil immersion can be used to reduce refraction at high magnification

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6
Q

What resolution does a microscope need to be in to distinguish if two cells are separate cells

A

0.2 um (with a long tail in the beginning of the u)

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7
Q

Define resolution

A

The ability to distinguish two adjacent points as distinct objects

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8
Q

What is resolution determined by

A

Resolution (resolving power) is determined by the wavelength of light used and the lens quality

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9
Q

What does it mean to have a smaller resolution value

A

It means the microscope is better (ex: the shorter the distance between two objects can be before they appear as a single object

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10
Q

What type of cells can be stained?

A

Live or unstained

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11
Q

What does staining improve?

A

Contrast

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12
Q

What type of stain is used on live cells

A

Vital stains

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13
Q

Why is a mordant used?

A

It may be used to fix the stain or coat the specimen

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14
Q

Stain molecules are usually ___?

A

Charged

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15
Q

Define fixation?

A

Tissue (cells) treated with a chemical that kills them and links the proteins together (preserves structures)

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16
Q

What type of stain is a gram stain

A

Differential

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17
Q

When is the gram stain done?

A

Often the first test done when identifying a bacteria culture

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18
Q

How important is the gram stain?

A

It is the most important prokaryote staining technique to date - modern and historic

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19
Q

What does the gram stain reveal?

A

It reveals two types of bacterial cell walls (gram - and gram +)

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20
Q

What chemical is used for primary stain?

A

Crystal Violet

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21
Q

What chemical is used for mordant?

A

Iodine

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22
Q

What chemical is used for decolourizing agent?

A

Alcohol-acetone

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23
Q

What chemical is used for counterstain?

A

Safranin

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24
Q

What colour are gram + and gram - stains in primary stain?

A

Gram +: Purple

Gram -: Purple

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25
Q

What colour do gram + and gram - stain with mordant

A

Gram +: Purple

Gram -: Purple

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26
Q

What colour do gram + and gram - stain with decolourizing agent?

A

Gram +: Purple

Gram -: White

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27
Q

What colour do gram + and gram - stain with counterstain?

A

Gram +: Purple

Gram -: Pink

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28
Q

Give an example of Gram + and what colour do they stain?

A

Staphylococcus aureus

They stain purple

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29
Q

Give an example of Gram - and what colour do they stain?

A

Echerichia coli

PInk

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30
Q

What do bacterial cell walls contain?

A

Mycolic Acid (waxy) and do not take up most stains

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31
Q

Name a bacteria you would stain using an acid-fast stain?

A

Mycobacterium tuberculosis

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32
Q

What is the other name for acid fast staining?

A

Ziel-Neelson Acid Fast Stain

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33
Q

What does the acid do in acid-fast staining?

A

Acid is used to decolourize

34
Q

What does the ‘fast’ do in acid-fast staining?

A

Fastness = stainis ‘locked in’

35
Q

How does acid-fast staining work?

A

It can be stained with concentrated dye + heat

- These organisms resist decolourization by dilute acid

36
Q

What is the primary stain in acid fast staining?

A

Heat + Carbol Fuchsin

37
Q

What is the decolourizing agent in acid-fast staining?

A

Acid alcohol

38
Q

What is the counterstain in acid-fast staining?

A

Methylene blue

39
Q

What colour do acid-fast and non-acid fast bacteria stain with primary stain?

A

Acid-Fast: Pink

Non-Acid-Fast: Pink

40
Q

What colour do acid-fast and non-acid fast bacteria stain with decolourizing agent?

A

Acid-Fast: Pink

Non-Acid-Fast: White

41
Q

What colour do acid-fast and non-acid fast bacteria stain with counterstain?

A

Acid-Fast: Pink

Non-Acid-Fast: Blue

42
Q

Describe the 3 steps of acid fast staining

A
  1. Stain (carbol fuschin) is driven into the cells with heat
  2. Cells are then decolourized with acid-alcohol
    - All cells (except acid fast bacteria) will decolourize
  3. Other cells are then counterstained with methylene blue
43
Q

What colour do mycobacterium tuberculosis stain?

A

Red/Pink

44
Q

What colour do you stain the endospore stain?

A

Red/Pink

45
Q

What type of stain is a capsule stain?

A

A negative stain

46
Q

What colour do capsule stains?

A

Purple with a white capsule around it

47
Q

What colour do flagellas stain?

A

Red/Pink

48
Q

What does a fluorescence microscope use to see structures and at what resolution?

A

Use florescent stains to illuminate specific cellular structures at high resolution

49
Q

What does a confocal microscope use to see things and at what depth?

A

Confocal microscopy used lasers to focus light or excite fluorescent dyes at precise depths

50
Q

What can you see under the fluorescent and confocal microscope?

A

Mucus and debris

- Mycobacterium tuberculosis

51
Q

Name the order of the electromagnetic spectrum from best resolution to worst

A
  • Gamma Rays
  • Electron Waves
  • X-rays
  • UV radiation
  • Visible light
  • Infrared radiation
  • Microwaves
  • Radio waves
52
Q

The colours of the rainbow consume what?

A

Visible light (400-700 nm)

53
Q

The Electron Microscope does what?

A

Forms images using electrons instead of light

54
Q

How are electrons focused in the electron microscope?

A

With magnets rather than glass lenses

55
Q

What does the Scanning Electron Microscope (SEM) do what to electrons?

A

Heavy metal coating reflects secondary electrons

56
Q

What bacteria can you see with a SEM?

A

Escherichia coli

57
Q

What animal can you see with a SEM?

A

Mosquito

58
Q

What does TEM stand for?

A

Transmission Electron Microscope

59
Q

Describe 3 things about the TEM

A
  • specimen must be extremely thin
  • electrons pass through the specimen
  • specimens are stained with heavy metals (eg. tungstun)
60
Q

What is the magnification of TEM?

A

10,000 - 100,000x

61
Q

What components of E.Coli can you see under the TEM microscope?

A
  • E. coli plasma membrane
  • Bdellovibrio
  • E. coli cell wall
62
Q

What are TEMS used to study?

A

Mainly used to study the internal structure of cells

63
Q

What type of bacteria can you see with a TEM?

A

Bacilllus coagulans

64
Q

What components of a Bacillus coagulans can you see with a TEM?

A
  • Capsule
  • Cell wall
  • Plasma membrane
  • Ribosomes
  • Nucleoid
    (you can not see the flagella, fimbriae, or bacterial chromosome)
65
Q

What is the illumination of a light microscopy?

A

Light

66
Q

What is the illumination of an electron microscopy?

A

Electrons

67
Q

What type of lense(s) do light microscopes have?

A

Glass/Plastic/Quartz

68
Q

What type of lens does electron microscope have?

A

None

69
Q

What type of view does the light microscope have?

A

Eye, camera

70
Q

What type of view does the electron microscope have?

A

screen, photograph paper

71
Q

Does the light microscope have colour?

A

Yes

72
Q

Does the electron microscope have colour?

A

No

73
Q

Are specimens alive in the light microscope?

A

They can be dead or alive

74
Q

Are specimens alive in the electron microscope?

A

They are dead

75
Q

What are the techniques for the light microscope?

A
  • Oil immersion
  • staining
  • fluorescence/confocal
76
Q

What are the techniques for the electron microscope?

A
  • SEM

- TEM

77
Q

What is the cost/ease of the light microscope?

A

Easy and cheap

78
Q

What the cost/ease of the electron microscope?

A

It is expensive and requires training

79
Q

What is the magnification of the light microscope?

A

100-1000x

80
Q

What is the magnification of the electron microscope?

A

SEM: up to 250,000
TEM: up to 1,000,000X