Professional Documents
Culture Documents
uk
YOUR NOTES
1.1 The Microscope in Cell Studies ⬇
CONTENTS
1.1.1 The Microscope in Cell Studies
In order to observe cellular material in more detail, specimens can be prepared for viewing
under a light microscope
The type of preparation that is appropriate is dependent on the cellular material that needs
to be viewed
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
YOUR NOTES
1.1 The Microscope in Cell Studies ⬇
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 magnification under which the observations shown by the drawing are made
must be recorded
No shading
The drawing should take up as much of the space on the page as possible
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
1.1 The Microscope in Cell Studies ⬇
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 The Microscope in Cell Studies ⬇
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
YOUR NOTES
1.1 The Microscope in Cell Studies ⬇
Worked Example
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
YOUR NOTES
1.1 The Microscope in Cell Studies ⬇
YOUR NOTES
1.1 The Microscope in Cell Studies ⬇
Worked Example
2000 / 1000 = 2, so the actual thickness of the leaf is 2 mm and the drawing thickness is 50 mm
So the magnification is x 25
YOUR NOTES
1.1 The Microscope in Cell Studies ⬇
An eyepiece graticule and stage micrometer are used to measure the size of the object
when viewed under a microscope
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
YOUR NOTES
1.1 The Microscope in Cell Studies ⬇
In the diagram, the stage micrometer has three lines each 10 µm apart
40 graticule divisions = 10 µm
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:
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 The Microscope in Cell Studies ⬇
To calculate the total magnification the magnification of the eyepiece lens and the
objective lens are multiplied together:
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 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
1.1 The Microscope in Cell Studies ⬇
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
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
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
1.1 The Microscope in Cell Studies ⬇
YOUR NOTES
1.1 The Microscope in Cell Studies ⬇
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
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
1.1 The Microscope in Cell Studies ⬇
Known values:
Image size: 3 mm
Step 2: Calculate the image size in the units asked for (micrometres)
1 mm = 1000 μm
3 mm = 3000 μm
Therefore, the average size of a red blood cell in this sample is 7.5 micrometres
YOUR NOTES
1.1 The Microscope in Cell Studies ⬇
YOUR NOTES
1.1 The Microscope in Cell Studies ⬇