2.1.3 Microscopy Flashcards

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1
Q

Define magnification

A

How many times bigger an object appears

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2
Q

Define resolution

A

The ability to distinguish two points next to each other

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3
Q

If the resolution of a microscopic image is too low, what can be said about the light/electrons used

A

The wavelength is too long

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4
Q

Magnification equation

A

Magnification = image size / actual size

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5
Q

1cm = ?mm

A

10

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6
Q

1mm = ?µm

A

1000

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7
Q

1µm = ?nm

A

1000

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8
Q

Measurement unit conversion image

A
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9
Q

How to measure an object using a microscope and graticule

A
  • Line up graticule with object
  • Count how many epu for the measurement
  • Calibrate graticule using stage micrometer - how many µm per epu
  • Multiply the number of epu by the calibration factor
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10
Q

Resolution of optical microscope

A

Low

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11
Q

Resolution of SEM

A

Medium

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12
Q

Resolution of TEM and ADVANTAGE

A
  • High
  • Allows organelles to be seen in greater detail
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13
Q

Magnification of optical microscope

A

Low

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14
Q

Magnification of TEM

A

High

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15
Q

Magnification of SEM

A

Medium

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16
Q

How does a optical microscope work

A

Light ray focused onto object then onto eye using glass lenses

17
Q

How does a SEM work

A
  • Electrons deflected/bounce off specimen
  • Foccused by electromagnets
18
Q

How does a TEM work

A
  • Electrons transmitted/pass through specimen
  • Foccused by electromagnets
19
Q

Primary use of optical microscope

A

Study living cells

20
Q

Primary use of SEM

A

View 3D image

21
Q

Primary use of TEM

A

View cell ultrastructure

22
Q

Are optical microscope images in colour

A

Yes

23
Q

Are TEM images in colour

A

No

24
Q

Are SEM images in colour

A

No

25
Q

Non-limitations to optical microscopes

A
  • Can be dead or alive
  • No vacuum
  • Not too thin
  • Stained with dye
  • No artefacts
  • Doesn’t take too long
26
Q

Limitations to SEM

A
  • Must be dead
  • In vacuum
  • Very thin specimin
  • Stained with metal
  • Artefacts may occur
  • Time consuming
27
Q

Limitations to TEM

A
  • Must be dead
  • Must be in vacuum
  • Specimin must be very thin
  • Stained with metal
  • Artefacts may occur
  • Time consuming
28
Q

In microscope images, if the resolution is high/low what can be said about the light or electrons wavelength

A
  • Low resolution = long wavelength
  • High resolution = short wavelength
29
Q

Describe how to extract chloroplast from a cell sample

A
  • Place sample in cold, pH buffered, isotonic solution
  • Homogenise in homogeniser
  • Place homogenate in test tube and spin in centrifuge
  • Remove nuclei pellet (supernatant) at bottom
  • Place homogenate in new test tube and spin at higher speed
  • Remove chloroplast supernatant
30
Q

When extracting organelles from cells during ultracentrifugation, what is the order of heaviest organelles, that must be removed

A
  • Nuclei
  • Chloroplast - only if plant cell
  • Mitochondria
31
Q

When extracting organelles from cells, why must the solution be cold

A

Reduce enzyme activity, so organelles aren’t digested

32
Q

When extracting organelles from cells, why must the solution be isotonic

A

Prevent water movement into organelles by osmosis - preventing lysis

33
Q

When extracting organelles from cells, why must the solution be pH buffered

A
  • Maintain pH
  • Prevent denaturations of proteins and enzymes