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Biology > M2: Microscopes & Staining > Flashcards

M2: Microscopes & Staining Flashcards

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1
Q

2.1.0 Foundations in Biology:

What’s Magnification?

I = AM

A

How much bigger an image is than the actual object

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2
Q

2.1.0 Foundations in Biology:

What’s meant by Resolution?

A

Distance between 2 points so they can’t be seen as one

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3
Q

2.1.0 Foundations in Biology: Light Microscope

What’s a Light Microscope?

A

Beam of light focused w lenses to produce an image

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4
Q

2.1.0 Foundations in Biology: Light Microscope

What are the Advantages of using a Light Microscope?

A
  • Easy to use
  • Cheaper
  • Studies living organisms
  • Coloured image
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5
Q

2.1.0 Foundations in Biology: Light Microscope

What are the Disadvantages of using a Light Microscope?

A
  • 200nm resolution
    ↳ long wavelength of light
  • x1500 max mag
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6
Q

2.1.0 Foundations in Biology: Laser Microscope

What’s a Laser Microscope?

A

Scans object point by point & computer assembles it into an image

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7
Q

2.1.0 Foundations in Biology: Laser Microscope

What are the Advantages of using a Laser Microscope?

A
  • Studies living organisms
  • Has depth selectivity
    ↳ focuses on structures at diff depths
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8
Q

2.1.0 Foundations in Biology: Laser Microscope

What are the Disadvantages of using a Laser Microscope?

A
  • More expensive
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9
Q

2.1.0 Foundations in Biology: TEM

What’s a TEM?

(Transmittion Electron Microscope)

A

Electromagnets used to focus beam of electrons through specimen

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10
Q

2.1.0 Foundations in Biology: TEM

What are the Advantages of using a TEM?

A
  • 0.1nm max resolution
  • x5,000,000 max mag
  • Not alive
    ↳ used for internal structures
  • 2D
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11
Q

2.1.0 Foundations in Biology: TEM

What are the Disadvantages of using a TEM?

A
  • Expensive
  • Training needed to use
  • Black & White image
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12
Q

2.1.0 Foundations in Biology: SEM

What’s a SEM?

(Scanning Electron Microscope)

A

Scans beams of electrons across specimen

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13
Q

2.1.0 Foundations in Biology: SEM

What are the Advantages of using a SEM?

A
  • 1nm max resolution
  • x500,000 max mag
  • Not alive
    ↳ surface of specimen
  • 3D
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14
Q

2.1.0 Foundations in Biology: SEM

What are the Disadvantages of using a SEM?

A
  • Expensive
  • Black & White
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15
Q

2.1.0 Foundations in Biology: Pre-prepared slides

What occurs in the Fixation stage?

A

Chemicals preserve material in life like condition

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16
Q

2.1.0 Foundations in Biology: Pre-prepapred slides

Why does the Fixation stage occur?

A

So specimen isn’t distorted

17
Q

2.1.0 Foundations in Biology: Pre-prepared slides

What occurs in the Dehydration stage?

A

Water is removed from specimen using ethanol

18
Q

2.1.0 Foundations in Biology: Pre-prepared slides

Why does the Dehydration stage occur??

A

Important for electron microscope
↳ water molecules deflect electron beam → blurs image

19
Q

2.1.0 Foundations in Biology: Pre-prepared slides

What occurs in the Embedding stage?

A

Supports the tissue in wax/resin
↳ so it can be cut into sections

20
Q

2.1.0 Foundations in Biology: Pre-prepared slides

Why does the Embedding stage occur?

A
  • Sectioning produces very thin slices of mounting
  • Sections are cut to make them a few um (light microscope) or nm(electron microscope)
21
Q

2.1.0 Foundations in Biology: Pre-prepared slides

Why is Staining used?

A

Cell material = transparent
* Staining used → increase contrast between different structures

22
Q

2.1.0 Foundations in Biology: Pre-prepared slides

Why is Mounting used?

A

Protects material so its suitable for viewing over a long period

23
Q

2.1.0 Foundations in Biology: Setting up a slide

How would you set up a Dry Mount?

A
  • Thin slice (sectioning) using a sharp knife
    ↳ increases resolution → light can pass through it
  • Place coverslip on top
24
Q

2.1.0 Foundations in Biology: Setting up a slide

How would you set up a Wet Mount?

A
  • Pipette small amounts of water onto the slide
  • Use tweezers to place specimen
  • Use a coverslip at an angle
    ↳ avoid trapping air bubbles
  • Add stain to one side of the coverslip
25
Q

2.1.0 Foundations in Biology: Components

What are Positive Charged Stains?

A

Stains negative charged materials in cytoplasm

26
Q

2.1.0 Foundations in Biology: Components

What are Positive Charged Stains used for?

A

Staining cell components (floating within the liquid)

27
Q

2.1.0 Foundations In Biology: Components

What are examples of Positive Charged Stains?

A
  • Methylene Blue
  • Crystal Violet
28
Q

2.1.0 Foundations in Biology: Background

What are Positively Charged Stains?

A

Stains negatively charged liquids
* Stains background not the specimen (liquid itself)

29
Q

2.1.0 Foundations in Biology: Background

What are Positively Charged Stains used for?

A

Stains background not the specimen (liquid itself)

30
Q

2.1.0 Foundations in Biology: Background

What are examples of Positively Charged Stains?

A
  • Nigrosin
  • Congo Red
31
Q

2.1.0 Foundations in Biology: Negatively Charged

What are Acidic Stains?

Eosin & Carbonfuchsin

A

Stains positively charged molecules e.g proteins

32
Q

2.1.0 Foundations in Biology: Negatively Charged

What are examples of Acidic Stains?

A
  • Eosin
  • Carbonfuchsin
33
Q

2.1.0 Foundations in Biology: Use in staining

What is the purpose of Crystal Violet?

A

Just stains cell purple/blue

34
Q

2.1.0 Foundations in Biology: Use in staining

What is the purpose of Iodine?

A

Ensures this colour is set

35
Q

2.1.0 Foundations in Biology: Use in staining

What is the purpose of Alcohol/Ethanol?

A

Rinses cell to ensure it stays purple/blue (if gram+) or turns colourless (if gram-)

36
Q

2.1.0 Foundations in Biology: Use in staining

What is the purpose of Safranin?

A

Stain where gram+ stays purple/blue and gram- appears pink/red

Biology (14 decks)

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