1.1.1 The Microscope in Cell Studies

CIE AS Biology (9700) exams from 2022 Revision Notes savemyexams.co.
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YOUR NOTES
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
1.1.1 THE MICROSCOPE IN CELL STUDIES

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
CIE AS Biology (9700) exams from 2022

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CIE AS Biology (9700) exams from 2022 Revision Notes savemyexams.co.uk
YOUR NOTES
Common microscope stains & uses table
Drawing Cells
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)

YOUR NOTES
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.

YOUR NOTES
1.1.2 MAGNIFICATION CALCULATIONS

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

YOUR NOTES
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:
The size of cells is typically measured using the micrometre (μm) scale, with cellular
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

YOUR NOTES
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)

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

YOUR NOTES
1.1.3 EYEPIECE GRATICULES & STAGE MICROMETERS

Eyepiece Graticules & Stage Micrometers
An eyepiece graticule and stage micrometer are used to measure the size of the object
when viewed under a microscope
Each microscope can vary slightly so needs to be calibrated when used
The calibration is done with a stage micrometer, this is a slide with a very accurate scale in
micrometres (µm), it is usually in 10 µm divisions, so 1 mm divided into 100 divisions
The eyepiece graticule is a disc placed in the eyepiece with 100 divisions, this has no scale
To know what the divisions equal at each magnification the eyepiece graticule is calibrated to
the stage micrometer at each magnification
Using stage micrometer & eyepiece graticule

YOUR NOTES
In the diagram, the stage micrometer has three lines each 10 µm apart
Each 10 µm division has 40 eyepiece graticule divisions
40 graticule divisions = 10 µm
1 graticule division = number of micrometres ÷ number of graticule division
1 graticule division = 10 ÷ 40 = 0.25 µm this is the magnification factor
The specimen slide would be used to replace the stage micrometer and the eyepiece
graticule at the same magnification would 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)
Exam Tip
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.

YOUR NOTES
1.1.4 RESOLUTION & MAGNIFICATION

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”

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

YOUR NOTES
Light v Electron Microscope Table

YOUR NOTES
1.1.5 CALCULATING ACTUAL SIZE

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.

YOUR NOTES
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
Therefore, the average size of a red blood cell in this sample is 7.5 micrometres

YOUR NOTES
Exam Question: Easy
Exam Question: Medium

YOUR NOTES
Exam Question: Hard
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CIE AS Biology (9700) exams from 2022 Revision Notes savemyexams.co.

1.1.2 Magniﬁcation Calculations

1.1.3 Eyepiece Graticules & Stage Micrometers

1.1.4 Resolution & Magniﬁcation

1.1.5 Calculating Actual Size

1.1.1 THE MICROSCOPE IN CELL STUDIES

Samples need to be thin enough to allow light to pass through

Slide preparation methods table

CIE AS Biology (9700) exams from 2022

Common microscope stains & uses table

The conventions are:

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)

Well-deﬁned structures should be drawn

The drawing should be made with proper proportions

CIE AS Biology (9700) exams from 2022

CIE AS Biology (9700) exams from 2022

1.1.2 MAGNIFICATION CALCULATIONS

Magniﬁcation is how many times bigger the image of a specimen observed is in

An equation triangle for calculating magniﬁcation

CIE AS Biology (9700) exams from 2022

An image of an animal cell is 30 mm in size and it has been magniﬁed by a factor of X

To ﬁnd the actual size of the cell:

Magniﬁcation does not have units

CIE AS Biology (9700) exams from 2022

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)

CIE AS Biology (9700) exams from 2022

Step 1: Check that units in magniﬁcation questions are the same

Remember that 1mm = 1000µm

Step 2: Calculate Magniﬁcation

Magniﬁcation = image size / actual size = 50 / 2 = 25

CIE AS Biology (9700) exams from 2022

1.1.3 EYEPIECE GRATICULES & STAGE MICROMETERS

Each microscope can vary slightly so needs to be calibrated when used

Using stage micrometer & eyepiece graticule

CIE AS Biology (9700) exams from 2022

Each 10 µm division has 40 eyepiece graticule divisions

1 graticule division = number of micrometres ÷ number of graticule division

1 graticule division = 10 ÷ 40 = 0.25 µm this is the magniﬁcation factor

graticule divisions x magniﬁcation factor = measurement (µm)

CIE AS Biology (9700) exams from 2022

1.1.4 RESOLUTION & MAGNIFICATION

A light microscope has two types of lens:

A series of (usually 3) objective lenses, each with a diﬀerent magniﬁcation

eyepiece lens magniﬁcation x objective lens magniﬁcation

The resolution of a light microscope is limited by the wavelength of light

As light passes through the specimen, it will be diﬀracted

CIE AS Biology (9700) exams from 2022

Comparison of the electron microscope & light microscope

The specimens can be living (and therefore can be moving), or dead

Due to the higher frequency of electron waves (a much shorter wavelength)

CIE AS Biology (9700) exams from 2022

Light v Electron Microscope Table

CIE AS Biology (9700) exams from 2022

1.1.5 CALCULATING ACTUAL SIZE

Worked example: Calculating the actual size of a specimen

CIE AS Biology (9700) exams from 2022

Eyepiece lens magniﬁcation: x10

Objective lens magniﬁcation: x40