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1.1.3Cell Level Systems

Electron Microscopes — OCR Gateway GCSE Combined Science A J250 - Biology Revision Notes & Topical Questions

5 min read 9 parts Lesson 4 of 13

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Part 1 of 7The need for greater resolving power

The light microscope is limited by the wavelength of visible light, so it cannot resolve objects smaller than about 0.2 micrometres.

Structures such as ribosomes, the internal membranes of mitochondria and the details of the nucleus are smaller than this.
Electron microscope
In the 1930s the electron microscope was invented.
It uses a beam of electrons instead of light.
Electrons have a much shorter wavelength than visible light,
  • which allows far greater resolution, and magnifications of more than ×1 000 000 are possible.

Part 2 of 7Two types

A transmission electron microscope (TEM) passes electrons through a very thin slice of specimen.

Two types
Denser parts absorb or scatter more electrons and appear darker,
  • giving a 2-D image of the internal structure of cells, such as the details of organelles.
Scanning electron microscope (SEM)
A scanning electron microscope (SEM) bounces electrons from the surface of the specimen, giving a detailed 3-D view of its surface.
In both cases, the electrons are focused using electromagnets, and the image is displayed on a screen or photographed,
  • producing a black and white image (an electron micrograph) that may be coloured by computer afterwards.

Part 3 of 7Comparison with the light microscope

FeatureLight microscopeElectron microscope
Radiation usedVisible lightBeam of electrons
Maximum magnificationAbout ×1500More than ×1 000 000
ResolutionAbout 0.2 micrometresAbout 0.1 nanometres (TEM)
SpecimenCan be livingDead, in a vacuum
ImageColour (with stains)Black and white
Cost and sizeCheap, portableVery expensive, large, needs training

The specimen must be dead for an electron microscope because the electrons only travel through a vacuum, and the preparation, which includes fixing, dehydrating and staining with heavy metals, can create artefacts: structures in the image that were not present in the living cell.

Part 4 of 7What electron microscopy revealed

Electron microscopes have allowed biologists to see sub-cellular structures in detail.

What electron microscopy revealed
They showed the internal folds of mitochondria (cristae) where respiration takes place, the stacks of membranes in chloroplasts, ribosomes, the nuclear envelope with its pores,
  • and the structure of bacteria and viruses.
They also showed the surface of structures such as pollen grains, insect eyes and cilia.
This understanding of the organelles helped to explain how each cell works, for instance how the folded inner membrane of a mitochondrion gives a large surface area for respiration.

Part 5 of 7Calculations with scale

For electron micrographs, the units are often micrometres or nanometres. 1 mm = 1000 µm and 1 µm = 1000 nm.

Calculations with scale
When a scale bar is given, measure the bar in the image, work out its real length,
  • and use it to calculate the magnification: magnification = measured length of scale bar / real length it represents.
Then use image size / magnification to find actual sizes.
Always write the units, and standard form is useful for very small values, for example 0.0000025 m is 2.5 × 10-6 m.

Part 6 of 7Worked example: reading a micrograph

Suppose a TEM image shows a chloroplast and a scale bar of 20 mm that represents 2 micrometres.

Worked example: reading a micrograph
The magnification is 20 mm / 0.002 mm = ×10 000.
  • If the chloroplast in the image is 50 mm long, then its actual length is 50 / 10 000 = 0.005 mm = 5 micrometres.
This kind of two-step calculation, finding the magnification from the scale bar and then using it, appears often.

When describing what you can see on a micrograph, name the organelles by their features: a mitochondrion appears as a small oval with folded inner membranes; a nucleus is a large round structure with a double membrane and pores; ribosomes are tiny dark dots; chloroplasts have stacks of membranes; and a cell wall appears as a thick line outside the membrane in plant cells.

Applications also extend beyond cellsSEM images are used to study the structure of insects, pollen and fibres, while TEM is used to study viruses and the details of the cell membrane.

Worked example: reading a micrograph
Both are used in medical diagnosis, for instance identifying a virus in a patient sample.
The limits to bear in mind are cost, specimen preparation and the risk of artefacts,
  • which is why electron microscopy results are checked against other methods.

Part 7 of 7Exam-style questions

  1. An electron micrograph shows a mitochondrion that measures 45 mm long. The magnification is ×30 000. Calculate the actual length in micrometres. [3]3 marks

  2. Explain why electron microscopes have been able to show more detail of cell structures than light microscopes. [3]3 marks

Key terms

TermMeaning
Electron microscopeA microscope that uses a beam of electrons to give high magnification and resolution.
TEMTransmission electron microscope, which shows thin sections in 2-D.
SEMScanning electron microscope, which shows the surface in 3-D.
ResolutionThe ability to see two close points as separate.
WavelengthThe distance between two successive peaks of a wave.
ArtefactA feature on an image created by the preparation, not present in the living specimen.
NanometreOne billionth of a metre (1 nm = 0.001 micrometre).

Avoid these mistakes

1
Saying electron microscopes have greater magnification but forgetting to say that the key advantage is resolution.
2
Saying electron microscopes can view living specimens. The specimen must be in a vacuum, so is dead.
3
Forgetting to convert units (mm to micrometres) in magnification questions.
4
Saying electron micrographs show true colours; they are black and white unless computer-coloured.

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