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Biology > Microscopy > Flashcards

Microscopy Flashcards

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1
Q

Describe, in principle, what a microscope does. (F)

A

An instrument which enables you to magnify an object

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2
Q

Name 4 different types of microscope. (F)

A
  • Light Microscope
  • Transmitting Electron Microscope (TEM)
  • Scanning Electron Microscope (SEM)
  • Laser Scanning Confocal Microscope (LSCM)
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3
Q

State what “SEM” and “TEM” are abbreviations for. (F)

A
  • Scanning Electron Microscope

- Transmitting Electron Microscope

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4
Q

Outline how an SEM works.

A
  • beam of electrons sent across surface of specimen

- reflected electrons are collected

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5
Q

Outline how a TEM works.

A
  • beam of electrons transmitted through a specimen

- focused to produce an image

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6
Q

Outline how a laser scanning confocal microscope works.

A
  • single spot of focused light (laser) moved across a specimen (point illumination)
  • causes fluorescence from components labelled with “dye”
  • emitted light from specimen filtered through pinhole aperture
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7
Q

Describe the use and properties of light microscopy. (F)

A

Use:

  • easily available
  • can be used out in field

Properties:

  • relatively cheap
  • can view living/dead specimens
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8
Q

Describe the use and properties of SEM. (F)

A

Use:
- can produce images with high magnifications and clear resolution

Properties:

  • dead specimens (vacuum)
  • can see more detail of cell ultrastructure
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9
Q

Describe the use and properties of TEM. (F)

A

Use:
- can produce images with high magnifications and clear resolution

Properties:

  • dead specimens (vacuum)
  • can see more detail of cell ultrastructure
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10
Q

Describe the use and properties of LSCM. (F)

A

Use:

  • diagnosis of diseases of the eye and endoscopic procedures
  • used for drug development
  • virtual biopsies

Properties:

  • living/dead specimen
  • can see distribution of molecules within cells
  • non-invasive
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11
Q

State the features of the images produced from light microscopes.

A
  • true colour
  • living/dead specimen
  • halide stains (?)
  • 2D image
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12
Q

State the features of the images produced from SEM.

A
  • false colour
  • dead specimen (vacuum)
  • heavy metal stains
  • high magnification and resolution
  • 3D image
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13
Q

State the features of the images produced from TEM.

A
  • false colour
  • dead specimen (vacuum)
  • heavy metal stains
  • high magnification and resolution
  • 2D image
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14
Q

State the features of the images produced from LSCM.

A
  • true colour
  • living/dead specimen
  • fluorescent dye
  • 2D/3D (can scan in layers)
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15
Q

Explain how to use a light microscope to view a specimen at low and high powers.

A
  • change the objective lens to a different power

- adjust resolution with coarse and fine focusing knob

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16
Q

Describe how to produce a temporary wet mount of living tissue.

A
  • suspend specimen in liquid (e.g. water or an immersion oil)
  • place a cover slip on from an angle
17
Q

Describe and explain the characteristics of a good slide preparation.

A
  • suitable stain so can differentiate between different parts of the sample
  • no air bubbles
  • evenly and well stained
  • thin specimen
  • no artefacts
18
Q

Explain why slide preparations need to be thin. (F)

A
  • so light can shine through

- details can be seen

19
Q

Explain how to use a stage micrometer to work out the distance represented by the small divisions in an eyepiece graticule under 3 different objective lenses. (F)

A

1 graticule division = number of micrometres/number of graticule divisions

20
Q

Explain how to use a stage micrometer and eye-piece graticule to add a scale bar to a drawing.

A
  • measure feature of speciment with eyepiece graticule
  • convert to micrometres using calibration done with stage micrometer
  • draw scale bar on drawing, matching the length of the feature you measured and label with micrometres
21
Q

Explain how to use a stage micrometer and eye-piece graticule to calculate the size of a specimen.

A

graticule divisions x magnification factor = measurement

22
Q

Describe how to choose an appropriate number of significant figures, or decimal places to present data.

A

Use the same decimal places/significant figures as all of the other data is given in

23
Q

Explain how an adjustment to the “plane of focus” can alter what is viewed within a cell. (F)

A

Because a cell is 3D, the shapes can look different from different angles i.e. the mitochondria is sausage shaped, but can appear spherical if viewed from the end.

24
Q

Explain how a tissue slice might be misleading due to the very thin nature of the slice.

A

Can be distorted.

25
Q

Explain why staining is useful for light microscopy. (F)

A

Provides contrast between different parts of the sample, so can be more easily identified.

26
Q

Describe the properties a stain needs to have to be useful for light microscopy. (F)

A
  • binds to the sample

- coloured

27
Q

Describe how to prepare a stained specimen for viewing under a light microscope.

A
  • place on a slide to air dry
  • heat-fix by passing through a flame
  • will adhere to slide and take up stain
28
Q

State the rules for biological drawings. (F)

A
  • title
  • magnification stated
  • sharp pencil
  • white, unlined paper
  • use as much of the paper as possible
  • smooth, continuous lines
  • do not shade
  • clearly defined structures
  • correct proportions
  • no arrow heads on label lines
  • label lines parallel to top of page and drawn with a ruler
29
Q

State the magnification formula and represent the magnification formula in a triangle diagram. (F)

A

magnification = size of image/actual size of object

30
Q

Explain how to use the magnification formula. (F)

A
  • rearrange so what you want to find is the subject of the equation
  • substitute values in and solve
31
Q

State the symbols used for millimetres, micrometres and nanometres. (F)

A

millimetres: mm
micrometres: μm
nanometres: nm

32
Q

Explain how to convert measurements from one unit into another.

A
  • find the difference between the units
  • if converting to a bigger unit, divide the measurement by the difference
  • if converting to a smaller unit, multiply the measurement by the difference
33
Q

Explain how to represent numbers in standard form.

A
  • number before the decimal place can be from 1 to 9
  • count how many places decimal place moved
  • if moved left, power of 10 is +ve
  • if moved right, power of 10 is -ve
34
Q

Define the terms “resolution”. (F)

A

The ability to see individual objects as separate entities.

35
Q

State the difference between magnification and resolution.

A

Magnification is how much the image has been enlarged, whereas resolution is the ability to see detail.

36
Q

State the resolution and useful maximum magnification of light microscopes.

A
  • 200nm

- x 2000

37
Q

State the resolution and useful maximum magnification of SEMs.

A
  • 3-10nm

- x 100 000

38
Q

State the resolution and useful maximum magnification of TEMs.

A
  • 0.5nm

- x 500 000

39
Q

Define the term “magnification” (F)

A

How many times larger the image is than the actual size of the object being viewed.

Biology (23 decks)

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