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YOUR NOTES
A Level Biology CIE
1.1 The Microscope in Cell Studies
CONTENTS
1.1.1 The Microscope in Cell Studies
1.1.2 Magnification Calculations
1.1.3 Eyepiece Graticules & Stage Micrometers
1.1.4 Resolution & Magnification
1.1.5 Calculating Actual Size
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1.1.1 The Microscope in Cell Studies YOUR NOTES
Microscope Slide Preparation
In order to observe cellular material in more detail, specimens can be prepared for viewing
under a light microscope
Samples need to be thin enough to allow light to pass through
The type of preparation that is appropriate is dependent on the cellular material that needs
to be viewed
Slide preparation methods table
Samples sometimes need to be stained, as the cytosol and other cell structures may be
transparent or difficult to distinguish
To stain a slide the sample needs to be first air-dried and then heated by passing it
through a Bunsen burner flame – this will allow the sample to be fixed to the slide and to
take up the stain
As with the type of preparation required, the type of stain used is dependent on what type
of specimen is being used
Common microscope stains & uses table
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Drawing Cells YOUR NOTES
To record the observations seen under the microscope (or from photomicrographs taken)
a labelled biological drawing is often made
Biological drawings are line pictures which show specific features that have been
observed when the specimen was viewed
There are a number of rules/conventions that are followed when making a biological
drawing
The conventions are:
The drawing must have a title
The magnification under which the observations shown by the drawing are made must
be recorded
A sharp HB pencil should be used (and a good eraser!)
Drawings should be on plain white paper
Lines should be clear, single lines (no thick shading)
No shading
The drawing should take up as much of the space on the page as possible
Well-defined structures should be drawn
The drawing should be made with proper proportions
Label lines should not cross or have arrowheads and should connect directly to the
part of the drawing being labelled
Label lines should be kept to one side of the drawing (in parallel to the top of the page)
and drawn with a ruler
Drawings of cells are typically made when visualizing cells at a higher magnification power,
whereas plan drawings are typically made of tissues viewed under lower magnifications
(individual cells are never drawn in a plan diagram)
Exam Tip
When producing a biological drawing, it is vital that you only ever draw what you see
and not what you think you see.To accurately reflect the size and proportions of
structures you see under the microscope, you should get used to using the
eyepiece graticule.
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1.1.2 Magnification Calculations YOUR NOTES
Magnification Calculations
Magnification is how many times bigger the image of a specimen observed is in
comparison to the actual (real-life) size of the specimen
The magnification (M) of an object can be calculated if both the size of the image (I), and
the actual size of the specimen (A), is known
An equation triangle for calculating magnification
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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The size of cells is typically measured using the micrometre (μm) scale, with cellular YOUR NOTES
structures measured in either micrometers (μm) or nanometers (nm)
When doing calculations all measurements must be in the same units. It is best to use the
smallest unit of measurement shown in the question
To convert units, multiply or divide depending if the units are increasing or decreasing
Magnification does not have units
Converting units of measurement
There are 1000 nanometers (nm) in a micrometre (µm)
There are 1000 micrometres (µm) in a millimetre (mm)
There are 1000 millimetres (mm) in a metre (m)
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YOUR NOTES
Worked Example
Step 1: Check that units in magnification questions are the same
Remember that 1mm = 1000µm
2000 / 1000 = 2, so the actual thickness of the leaf is 2 mm and the drawing thickness is 50 mm
Step 2: Calculate Magnification
Magnification = image size / actual size = 50 / 2 = 25
So the magnification is x 25
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1.1.3 Eyepiece Graticules & Stage Micrometers YOUR NOTES
Eyepiece Graticules & Stage Micrometers
An eyepiece graticule and stage micrometer are used to measure the size of the object
when viewed under a microscope
The type of microscope and magnification used can vary signficantly so the eyepiece
graticule needs to be calibrated each time when measuring objects
The calibration is done using a stage micrometer, this is a slide with a very accurate known
scale in micrometres (µm)
The eyepiece graticule is a disc placed in the eyepiece with 100 divisions, this has no scale
To know what the graticule divisions equal at each magnification the eyepiece graticule is
calibrated to the stage micrometer at each magnification
Using stage micrometer & eyepiece graticule
A stage micrometer alongside an eyepiece graticule.
In the diagram, the stage micrometer has three lines each 100 µm (0.1 mm) apart
Each 100 µm division has 40 eyepiece graticule divisions
40 graticule divisions = 100 µm
1 graticule division = number of micrometres ÷ number of graticule division
1 graticule division = 100 ÷ 40 = 2.5 µm this is the magnification factor
The calibrated eyepiece graticule can be used to measure the length of the object
The number of graticule divisions can then be multiplied by the magnification factor:
graticule divisions x magnification factor = measurement (µm)
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Exam Tip YOUR NOTES
The calculations involving stage micrometers and eyepiece graticules are often
seen in exam questions, so make sure that you are comfortable with how to calibrate
the graticule and calculate the length of an object on the slide.
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1.1.4 Resolution & Magnification YOUR NOTES
Resolution & Magnification
Magnification
Magnification is how many times bigger the image of a specimen observed is in compared
to the actual (real-life) size of the specimen
A light microscope has two types of lens:
An eyepiece lens, which often has a magnification of x10
A series of (usually 3) objective lenses, each with a different magnification
To calculate the total magnification the magnification of the eyepiece lens and the
objective lens are multiplied together:
eyepiece lens magnification x objective lens magnification = total magnification
Resolution
Resolution is the ability to distinguish between two separate points
If two separate points cannot be resolved, they will be observed as one point
The resolution of a light microscope is limited by the wavelength of light
As light passes through the specimen, it will be diffracted
The longer the wavelength of light, the more it is diffracted and the more that this
diffraction will overlap as the points get closer together
Electron microscopes have a much higher resolution and magnification than a light
microscope as electrons have a much smaller wavelength than visible light
This means that they can be much closer before the diffracted beams overlap
The concept of resolution is why the phospholipid bilayer structure of the cell membrane
cannot be observed under a light microscope
The width of the phospholipid bilayer is about 10nm
The maximum resolution of a light microscope is 200nm (half the smallest wavelength
of visible light, 400nm)
Any points that are separated by a distance less than 200nm (such as the 10nm
phospholipid bilayer) cannot be resolved by a light microscope and therefore will not
be distinguishable as “separate”
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YOUR NOTES
The resolving power of an electron microscope is much greater than that of the light
microscope, as structures much smaller than the wavelength of light will interfere with a
beam of electrons
Comparison of the electron microscope & light microscope
Light microscopes are used for specimens above 200 nm
Light microscopes shine light through the specimen, this light is then passed through
an objective lens (which can be changed) and an eyepiece lens (x10) which magnify
the specimen to give an image that can be seen by the naked eye
The specimens can be living (and therefore can be moving), or dead
Light microscopes are useful for looking at whole cells, small plant and animal
organisms, tissues within organs such as in leaves or skin
Electron microscopes, both scanning and transmission, are used for specimens above
0.5 nm
Electron microscopes fire a beam of electrons at the specimen either a broad static
beam (transmission) or a small beam that moves across the specimen (scanning)
The electrons are picked up by an electromagnetic lens which then shows the image
Due to the higher frequency of electron waves (a much shorter wavelength)
compared to visible light, the magnification and resolution of an electron microscope
is much better than a light microscope
Electron microscopes are useful for looking at organelles, viruses and DNA as well as
looking at whole cells in more detail
Electron microscopy requires the specimen to be dead however this can provide a
snapshot in time of what is occurring in a cell eg. DNA can be seen replicating and
chromosome position within the stages of mitosis are visible
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Light v Electron Microscope Table YOUR NOTES
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1.1.5 Calculating Actual Size YOUR NOTES
Calculating Actual Size
When investigating the size of organisms and biological structures you will use a
microscope of a specific magnification to produce an image
Photomicrographs are images obtained from a light microscope, these are used for
specimens above 200 nm (a bacteria cell is about 1000 nm)
Electron micrographs are images obtained from electron microscopes, both scanning
and transmission, these are used for specimens above 0.5 nm
Electron microscopes are useful for looking at organelles and biological molecules, eg.
DNA can be seen replicating
To better understand the images we produce using microscopes we need to know the
actual size of the specimen
Worked example: Calculating the actual size of a specimen
A scientist looks at a sample of red blood cells under a light microscope.
The eyepiece lens of the microscope has a magnification of x10 and an objective lens of
x40 was used to view the blood cells. The scientist takes a photomicrograph of the blood
cells, in which the average size of each cell is 3 mm.
What is the average size of the red blood cells in the sample? Give your answer in
micrometres.
Known values:
Eyepiece lens magnification: x10
Objective lens magnification: x40
Image size: 3 mm
Step 1: Calculate the total magnification of the specimen
eyepiece lens magnification x objective lens magnification
= total magnification
x10 x x40 = x400
Step 2: Calculate the image size in the units asked for (micrometres)
1 mm = 1000 μm
3 mm = 3000 μm
Step 3: Calculate the actual size of the red blood cell
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Therefore, the average size of a red blood cell in this sample is 7.5 micrometres YOUR NOTES
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