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Microscopes
GCSE

A-level

You may have used light

microscopes to look at the
general structure of cells and
can identify larger organelles
such as the nucleus.

You will need to know the main features and

limitations of an optical microscope.

You will know that to see internal

structures and to identify roles
within the cell, you need to use
an electron microscope.

You should be able to identify the differences

between optical and electron microscopes, including
scanning and transmission electron microscopes.
You will need to use simple equations to work out
the size and magnification of a specimen.

Using microscopes
Microscopes are used to observe and magnify cells. They enable us to have a clear, detailed
understanding of the different structures found within cells. There are two key features that
microscopes provide:
1

Magnification this is the extent to which the

image is larger than the object being viewed; it is
simply the factor by which the image is enlarged
compared to the original object.

magnification

Important terms to remember:

Magnification  enlargement
Resolution  detail

size of image
_______________
actual size

It is important that the same units are used for both image and object. The size of the cell
or organelle is usually measured in millimetres (mm) and will generally need to be converted
to micrometres (m) by multiplying this number by 1,000.
2

Resolution this is the ability of a microscope to distinguish between structures that are
very close together. The resolving power of a microscope is the minimum distance apart
that two objects can be for them to appear as separate items.
eye

Light (optical) microscopes

A light microscope uses lenses to focus rays of light through the specimen.
The light rays are then passed through a series of lenses that are responsible
for both the magnification and resolution. Different parts of the specimen
absorb varying amounts and wavelengths of light. Light that is not absorbed is
transmitted to the eye through the objective lens and eyepiece.
(NB: A light microscope and an optical microscope are the same.)

nal image
eyepiece

projector lens

objective

A light microscope has a resolution of approximately 0.2m

specimen

Magnification is up to  1,500

condenser lens

For example, if objective lens   40 and eyepiece   10, then

overall magnification  40  10   400

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Microscopes

The electron microscope uses beams of electrons instead of light rays. The beams of electrons
are bent and focused by electromagnetic lenses. Electron beams have a much shorter
wavelength and pass through objects that are very close together, therefore providing a much
better resolution. Electron microscopes also have a greater magnification of  500,000.
electron gun

anode
condenser
specimen
objective aperture lens
intermediate lens

projector lens

1. Transmission Electron Microscope (TEM)

The beams of electrons produced by the electron
gun are transmitted through the material. The
section of the specimen being investigated has
to be extremely thin, as electrons are much more
easily blocked than light. Denser parts of the
specimen absorb more electrons and appear
darker on the image produced. Other parts of the
image allow the electrons to pass through and
hit the screen, which then fluoresces, and these
parts appear bright. The flat, two-dimensional
image produced is called a micrograph.

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Electron microscopes

The preparation involves using chemicals to fix

the specimen, which may create artefacts. These
are structures not originally present or that alter
the specimen from its original condition.
uorescent screen

2. Scanning Electron Microscope (SEM)

The beams of electrons are bounced off the surface of the specimen and create an image from the
electrons that are reflected from the surface. The specimen used for SEM can be much thicker than that
used for TEM, as there is no need for the electrons to penetrate through. The electrons are scattered
by the specimen and as the scattering depends on the contours of the specimen surface, a threedimensional image is produced.

Limitations of electron microscopy

Although electron microscopes produce a higher magnification and greater resolution, there are a
number of limitations that need to be taken into account:
i.

There must be a vacuum, as electrons can be scattered in all directions by air

molecules.
ii. Specimens are always dead.
iii. Tissue being observed has to undergo several stages of preparation, and each stage
may alter the real appearance.
iv. Preparation may introduce artefacts.

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Microscopes
What are the differences between light and electron
microscopes?
The following table summarises the main differences between the two types of microscope.
Feature

Light microscope

Electron microscope

Radiation

Light rays

Beams of electrons

Focusing

Glass lenses objective lens

and eyepiece

Electromagnetic lenses

Magnification

 1,500

 500,000

Resolution

Low 0.2 m

High 0.0001m

Preparation of specimen

Lower chance of producing

artefacts

Greater chance of producing

artefacts

Specimen

Dead or alive, as no vacuum

Dead, as vacuum used

Image

Viewed through eyepiece

Viewed on fluorescent screen

Equipment cost

Relatively cheap

Very expensive

Working out the size and magnification of a specimen

Calculating the actual size
If the size of an image and the magnification is known, then the actual size of the specimen can be
calculated using the following equation:

actual size

size of the image

magnification

Remember that the size of the image is usually measured in mm and should be converted into m.
Worked example
cristae

inner
membrane

outer
membrane

matrix

If a mitochondria was magnified by  5,000, measure the diameter from point X to point Y and
calculate the actual size.
Size of the image from X to Y  50mm
Actual size  50  1,000
5,000
Answer  10m

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Microscopes

Use the following equation to work out the magnification:

magnification = size of the image (mm 1,000 to convert to m) actual size
I = image (usually in mm)

I
A

A = actual size (usually in +m)

M = magnification

Questions
1

What is meant by magnification?

Define the term resolution, as used in microscopy.

Suggest two reasons why structures seen in electron microscopes might not
accurately represent those in a living cell?

Describe the difference between SEM and TEM.

If the diameter of cell is 35mm and magnification is  500, what is the actual size?

If the size of an image is 75mm and the actual size is 5m, what is the magnification?

What is the advantage of using an optical microscope?

What is the magnification of an image of a chloroplast if the actual size is 8m and the
image is measured at 24mm?

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Calculating the magnification

Taking it Further
Now make more detailed notes about microscopes. Make a table of comparisons between optical
and electron microscopes, and a summary of the limitations of electron microscopes.
The questions below can help you to structure and organise your notes. Use your course text
book to add further information.
Name the different types of microscopes.
Which type of microscope has a better resolution?
What is the formula for calculating actual size?
Why are images obtained using electron microscopes not a true reflection of the specimen?
How can you measure magnification from a drawing provided?
List four differences between light microscopes and electron microscopes.
In a light microscope, if the eyepiece is  10 and the objective lens is  4, what is the
magnification?
What is the maximum magnification of optical microscopes used by A-level Biology
students in the laboratory?

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