1.

Cell Structure
1.1 The Microscope
in Cell Studies
Measuring size and calculating magnification
Do now: 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?
 There are 1000 nanometers (nm) in a micrometre (µm)

Converting units
There are 1000 micrometres (µm) in a millimetre (mm)
There are 1000 millimetres (mm) in a metre (m)

• 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
Using eyepiece graticules and stage micrometers
• Used to measure the size of the object when viewed under a microscope
• Eyepiece graticule must 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 eyepiece graticules and stage micrometers
• 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)
Do Now
 Work through the ‘Calculating Magnification’ examples on pg. 12
&13 of your CB and complete WB Ex. 1.5 Pg. 9

Homework - WB Ex.1.1 pg. 1 due next lesson

 Practical Work – 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
 Stains
• 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
Drawing Cells
• 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, but not extend over any writing or the page
• 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
Resolution and 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 and 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
Same magnification, different resolution
The electromagnetic spectrum CB Pg.
14-17
 Resolution is linked to light wavelength
 Visible light wavelength ranges from 400nm-700nm
 General rule: limit of resolution is about half the wavelength of the radiation used to view
the specimen
 What would the ‘best’ resolution obtained from a light microscope be?
The Electron Microscope
 Electron microscopes are used for specimens above 0.5 nm
 Electron microscopes fire a beam of electrons at the specimen. The electrons are picked up by an
electromagnetic lens which then shows the image
 Due to the higher frequency of electron waves (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
Do Now
1.2 Cells as the basic units of
living organisms
Ultrastructure of a typical eukaryote cell
Do Now
 Using your CB and various other resources, complete the self-study
of the organelles and cellular structures found in eukaryote cells.
 Complete the table as evidence of your self-study
 Structure of animal and plant cells
• The only structures found in animal cells but not plant cells are the centrioles and microvilli
• Plant cells also have additional structures: the cellulose cell wall, large
permanent vacuoles and chloroplasts
The ultrastructure of an animal cell shows a densely packed cell – the ER and RER and ribosomes form extensive networks
throughout the cell in reality
 The role of ATP
•All organisms require a constant supply of energy to maintain their cells and stay alive
•This energy is required:
• In anabolic reactions – building larger molecules from smaller molecules
• To move substances across the cell membrane (active transport) or to move substances within the
cell
• In animals, energy is required:
•For muscle contraction – to coordinate movement at the whole-organism level
•In the conduction of nerve impulses, as well as many other cellular processes
•In all known forms of life, ATP from respiration is used to transfer energy in all energy-requiring
processes in cells
•This is why ATP is known as the universal energy currency
•Adenosine Triphosphate (ATP) is a nucleotide (nitrogenous base + phosphate group + sugar)
• The monomers of DNA and RNA are also nucleotides
 General structure of a prokaryote

• unicellular
• generally 1–5 μm diameter
• peptidoglycan cell walls
• circular DNA
• 70S ribosomes
• absence of organelles surrounded by double membranes
E. coli ultrastructure
Draw the ultrastructure of E.coli,
including the cell wall, pili, flagella,
plasma membrane, cytoplasm, 70s
ribosomes, and nucleoid with naked
DNA.
Prokaryotes vs Eukaryotes
Prokaryotes have a cellular structure distinct from eukaryotes:
• Their genetic material is not packaged within a membrane-bound nucleus and is
usually circular (eukaryotic genetic material is packaged as linear chromosomes)
• Prokaryotes lack membrane-bound organelles
• They are many (100s/1000s) of times smaller than eukaryotic cells
• Their ribosomes are structurally smaller (70 S) in comparison to those found in
eukaryotic cells (80 S)
 Mnemonic:  DORA

 Prokaryotic cells and

eukaryotic cells differ in a
number of key features,
including:
• DNA (composition and
structure)
• Organelles (types present and
relative sizes)
• Reproduction (mode differs
according to chromosome
structure)
• Average size (exceptions may
exist)
Viruses
 Non-cellular, infectious particles
 Simple in structure
 Smaller than prokaryotes (20-300nm)
 They comprise of:
 Nucleic acid core (can be DNA or RNA; double or single stranded)
 Protein coat called CAPSID
 May have an outer envelope made of phospholipids
 All viruses are parasitic and can only reproduce by infecting living cells and using the host
cells ribosomes to produce new viral particles
Cell fractionation (differential
centrifugation)
Cell fractionation or differential centrifugation is a technique which separates organelles
according to their density - you might want to do this if you want to visualise certain
organelles under the microscope separately.
It involves bursting the cell surface membrane to release the organelles and spinning the
cell solution at really high speeds.

https://www.youtube.com/watch?v=_Y6Bzgkm6x8
Homogenisation
 First step of cell fractionation is homogenisation.
 This is where you break apart the plasma membrane to release the organelles.
 This can be done by vibrating the cells or by breaking them apart in a blender.
 It is important that this cells are placed into a solution which is ice-cold, isotonic and
buffered.
 the homogenised solution is filtered to remove any tissue debris.
 The organelles are small enough to pass through the holes of the filter paper so will be
present in the filtrate.
Ultracentrifugation
 The filtrate is spun at increasing speeds.
 The heaviest organelles will sink to the bottom of the test-tube, forming a
pellet.
 We can transfer the remaining solution (the supernatant) to a separate test
tube, which will be spun at a slightly higher speed.
 This is repeated until you obtain the organelle that you want.
 Remember that the organelles will be separated from the solution from the
heaviest to the lightest.
 Nuclei will come out of the solution first, followed by mitochondria, then
lysosomes, then the endoplasmic reticulum. Ribosomes will be the last
organelles to form a pellet, since these are the lightest organelles in a cell.
CB Exam Style Questions Pg. 36
You must complete the following questions:
 4, 6, 9c&d, 10