22/03/2025, 20:15 Light Microscopes | OCR GCSE Biology A (Gatew ay) Revision Notes 2016

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Light Microscopes
Author Last updated
Phil 20 November 2024

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Microscope Equipment
Many biological structures are too small to be seen by
the naked eye
Optical microscopes are an invaluable tool for scientists
as they allow for tissues, cells and organelles to be seen
and studied
Light is directed through a thin layer of biological
material (containing the tissue(s), cell(s) or organelle(s) to
be observed) that is supported on a glass slide
This light is focused through several lenses so that an
image is visible through the eyepiece

Apparatus
The key components of an optical microscope you will
need to use are:
The eyepiece lens
The objective lenses
The stage
The light source
The coarse and fine focus
Other apparatus used:
Forceps
Scissors
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22/03/2025, 20:15 Light Microscopes | OCR GCSE Biology A (Gatew ay) Revision Notes 2016

Scalpel
Coverslip Try 7 days free
Slides
Pipette

The components of an optical microscope

Calibration, Sample Prep & Setup

Preparing specimens & samples
Specimens must be prepared on a microscope slide to
be observed under a light microscope
This must be done carefully to avoid damaging the
biological specimen and the structures within it
The most common specimens to observe under a light
microscope are cheek cells (animal cells) and onion cells
(plant cells)
Preparing a slide using a liquid specimen:
Add a few drops of the sample to the slide using a
pipette
Cover the liquid/smear with a coverslip and gently
press down to remove air bubbles
Wear gloves to ensure there is no cross-
contamination of foreign cells
Preparing a slide using a solid specimen:
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22/03/2025, 20:15 Light Microscopes | OCR GCSE Biology A (Gatew ay) Revision Notes 2016

Use scissors to cut a small sample of the tissue

Peel away or cut a very thin layer of cellsTry
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tissue sample to be placed on the slide (using a
scalpel or forceps)
Some tissue samples need to be treated with
chemicals to kill/make the tissue rigid
Gently place a coverslip on top and press down to
remove any air bubbles
A stain may be required to make the structures visible
depending on the type of tissue being examined.
Commonly used stains include methylene blue
to stain cheek cells and iodine to stain onion cells
Take care when using sharp objects and wear gloves
to prevent the stain from dying your skin
Preventing the dehydration of tissue:
The thin layers of material placed on slides can dry up
rapidly
Adding a drop of water to the specimen (beneath the
coverslip) can prevent the cells from being damaged
by dehydration

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Care must be taken to avoid smudging the glass slide or

trapping air bubbles under the coverslip
Viewing the specimen
When using an optical microscope always start with the
low power objective lens:
It is easier to find what you are looking for in the field
of view
This helps to prevent damage to the lens or coverslip
in case the stage has been raised too high
Unclear or blurry images:
Switch to the lower power objective lens and try
using the coarse focus to get a clearer image
Consider whether the specimen sample is thin
enough for light to pass through to see the structures
clearly

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22/03/2025, 20:15 Light Microscopes | OCR GCSE Biology A (Gatew ay) Revision Notes 2016

There could be cross-contamination with foreign

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A calibrated graticule must be used to

take measurements of cells
A graticule is a small disc that has an engraved scale. It
can be placed into the eyepiece of a microscope to act
as a ruler in the field of view
As a graticule has no fixed units it must be calibrated for
the objective lens that is in use. This is done by using a
scale engraved on a microscope slide (a stage
micrometer)
By using the two scales together the number of
micrometers each graticule unit is worth can be worked
out
After this is known the graticule can be used as a ruler in
the field of view

The stage micrometer scale is used to find out how many

micrometers each graticule unit represents
Limitations of microscopy
The size of cells or structures of tissues may appear
inconsistent in different specimen slides
Cell structures are 3D and the different tissue
samples will have been cut at different
planes resulting in inconsistencies when viewed on
a 2D slide

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22/03/2025, 20:15 Light Microscopes | OCR GCSE Biology A (Gatew ay) Revision Notes 2016

Optical microscopes do not have the same magnification

power as other types of microscopes and soTry
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some structures that cannot be seen
The treatment of specimens when preparing slides could
alter the structure of cells

Using units in microscopy

You may be given a question in your Biology exam where
the measurements for a magnification calculation have
different units
You need to ensure that you convert them both into the
same unit before proceeding with the calculation (usually
to calculate the magnification)
Remember the following to help you convert between
mm (millimetres), µm (micrometres) and nm
(nanometres):

Converting between mm (millimetres), µm (micrometres)

and nm (nanometres)
If you are given a question with two different units in it,
make sure you make a conversion so that both
measurements have the same unit before doing your
calculation

Calculation of Magnification
Higher Tier Only
Magnification is calculated using the following equation:

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22/03/2025, 20:15 Light Microscopes | OCR GCSE Biology A (Gatew ay) Revision Notes 2016

Magnification = Drawing size ÷ Actual size

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One way to remember the equation is using an equation
triangle:

An equation triangle for calculating magnification

Rearranging the equation to find things other than the
magnification becomes easy when you remember the
triangle – whatever you are trying to find, place your
finger over it and whatever is left is what you do, so:
Magnification = image size ÷ actual size
Actual size = image size ÷ magnification
Image size = actual size × magnification
Remember magnification does not have any units and is
just written as ‘X 10’ or ‘X 5000’

Worked Example
An image of an animal cell is 30 mm in size and it has been
magnified by a factor of X 3000. What is the actual size of the
cell?
To find the actual size of the cell:

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Worked example using the equation triangle for

magnification

You may also be asked to calculate the total

magnification of a light microscope if given the
magnification of the eyepiece lens and the magnification
of the objective lens
As these are two separate parts of a light microscope,
each with its own magnifying power, you can simply
multiply the two values to calculate the total
magnification:
Magnification of light microscope = Magnification of
eyepiece lens × Magnification of objective lens

Standard form
When doing calculations and unit conversions, it is
common to come across very big or very small numbers
Standard form can be useful when working with these
numbers
Standard form is a way of writing very big and very small
numbers using powers of 10

How to use standard form

Using standard form, numbers are always written as
follows: a × 10n
The rules:
1 ≤ a < 10 (the number 'a' must always be between 1
and 10)

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22/03/2025, 20:15 Light Microscopes | OCR GCSE Biology A (Gatew ay) Revision Notes 2016

n > 0 for LARGE numbers ('n' = how many times 'a' is

multiplied by 10) Try 7 days free
n < 0 for SMALL numbers ('n' = how many times 'a' is
divided by 10)

Using standard form to convert between

units
For example, you can write 1 metre in millimetres using
standard form:
1 m = 1000 mm
So, 1 m = 1 mm × 1000
So, 1 m = 1 mm × 10 × 10 × 10
So, as we had to multiply 1 mm by 10 three times to
get 1 m, we write this as:
1 m = 1 × 103 mm
Writing 1 millimetre in metres using standard form is also
possible and is just the opposite:
1 mm = 0.001 m
So, 1 mm = 1 m ÷ 1000
So, 1 mm = 1 m ÷ 10 ÷ 10 ÷ 10
So, as we had to divide 1 m by 10 three times to get 1
mm, we write this as:
1 mm = 1 × 10-3 m
Exactly the same process can be used if you needed to
convert micrometres into millimetres. For example:
1 µm = 0.001 mm
So, 1 µm = 1 mm ÷ 1000
So, 1 µm = 1 mm ÷ 10 ÷ 10 ÷ 10
So, as we had to divide 1 mm by 10 three times to get
1 µm, we write this as:
1 µm = 1 × 10-3 mm

Examples of using standard form in

conversion calculations
You could be asked to state 45 centimetres in millimetres
using standard form:
1 cm = 10 mm
So, 45 cm = 450 mm
So, 45 cm = 4.5 mm × 10 × 10
So, as we had to multiply 4.5 mm by 10 two times to
get 45 cm, we write this as:
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22/03/2025, 20:15 Light Microscopes | OCR GCSE Biology A (Gatew ay) Revision Notes 2016

45 cm = 4.5 × 102 mm
You could also be asked to state 250 micrometres in free
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millimetres using standard form:
1 µm = 0.001 mm
So, 250 µm = 0.25 mm
So, 25 µm = 2.5 mm ÷ 10
So, as we had to divide 2.5 mm by 10 just once to get
250 µm, we write this as:
250 µm = 2.5 × 10-1 mm

Worked Example

Answer:

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22/03/2025, 20:15 Light Microscopes | OCR GCSE Biology A (Gatew ay) Revision Notes 2016

Step One: Convert units

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Remember that 1 mm = 1000 µm
So to get from µm to mm you need to divide by 1000
Step Two: Calculate the thickness of the leaf in mm
2000 ÷ 1000 = 2, so the actual thickness of the leaf is 2 mm
and the drawing thickness is 50 mm
Step Three: Put these values into the equation for
calculating magnification
Magnification = image size ÷ actual size = 50 ÷ 2 = 25
So the magnification is x 25

Examiner Tips and Tricks

It is easy to make silly mistakes with magnification
calculations. To ensure you do not lose marks in the exam:
Always look at the units that have been given in the
question – if you are asked to measure something, most
often you will be expected to measure it in millimetres
NOT in centimetres – double-check the question to see!
Learn the equation triangle for magnification and always
write it down when you are doing a calculation –
examiners like to see this!

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