LEARNING OUTCOMES

Define resolution and magnification

and explain the differences between
these terms, with reference to light
microscopy and electron microscopy
EARLY MICROSCOPES
2 types of microscopes:
Light microscope
Parts of a Compound Microscope
 Lenses:

Ocular Lens: eyepiece lens

Objective Lens: can be low, medium or high power

Look at magnification on lens
Lower power is smaller in size
• Arm: connects stage and body
tube

• Stage: platform with opening

over which a specimen is placed
(clips to hold slide)

• Base: supports microscope

 Letting in Light:

• Diaphragm:
regulates
amount of
light
• Eyepiece (ocular): part you look
through, holds ocular lens,
magnifies 10x

• Body tube: connects eyepiece &

objective lenses

• Nosepiece: holds objective lenses

(can be turned)

• Objective Lens: magnifies image,

can be low, medium, high power
Focusing:

Coarse Adjustment Knob:

use on low power only!!

Fine Adjustment Knob:

once low power is focused
switch to high power and use
fine adjustment.
Electron microscope
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 WHAT MAKES UP A GOOD MICROSCOPE?

Magnification = the number of times larger an image is

compared with the real size of the object

Magnification = Image size

Actual size
 WHAT MAKES UP A GOOD MICROSCOPE?
Resolution = ability to distinguish between two separate
points
Therefore, the higher the resolution, the more details you
can view under microscope
Visible lights is part of a spectrum of
electromagnetic energy radiating from the sun.
Different wavelengths of visible light are seen by
the human eye as different colors.

Gamma Micro- Radio

X-rays UV Infrared
rays waves waves

Visible light

Wavelength (nm)
 ELECTROMAGNETIC SPECTRUM
• Light travels in various wavelengths, λ known as
electromagnetic spectrum
• The longer the waves, the lower the frequency,
energy decreases
Why are plants green?

Transmitted light
 HOW is Resolution linked to light?
• Limit of resolution = about 1/2
wavelength of the radiation
used to view it
• Therefore, a microscope’s best
resolution is 200nm because
the shortest wavelength of
visible light is 400nm.
• Therefore, any object smaller
than 200nm cannot be
observed , does not interfere
with wavelength.
• Transparent objects are not
visible and needs to be stained
for viewing.
 What’s my actual size?

Magnification = 500X
 What’s my actual size? (cont’d)
This drawing of a mitochondrion has
been magnified 100 000x . What is
the actual size (in μm)?

This is a drawing of a chloroplast

(100 000x). What is the actual size
(in μm)?
 Electron microscope (EM)
Light microscope (LM)
Transmission EM Scanning EM
(TEM) (SEM)
Source Light Electron
Samples Live Dead
Resolution &
Lower (200nm) Higher (0.5 nm) Higher (3nm)
magnification

Electron beam Electron beam

Mechanism passes through irradiate surface
sample of object

• Can see living cells in

Advantages colours Higher resolution
• Cheaper
Only samples above • Samples will only appear in black
Disadvantages 200 nm can be and white
observed • More expensive
Check point question:
State the advantages of using the light microscope,
rather than using the electron microscope in
studies of plant and animal cells. [2]
• Can observe living tissues; A observing processes;
• Ref. portability; smaller in size, easy to move, no requirement
for special room;
• Ease of use; slide preparation easier, takes less time
• See (actual/natural/real-life) colour;
• Ref. to differential staining/staining particular types of tissues;
• Lower cost of purchase/maintenance/running/AW [max2]
LEARNING OBJECTIVE:
Describe and interpret photomicrographs,
electron micrographs and drawings of
typical plant and animal cells

Compare the structure of typical plant and

animal cells
 THE CELL?

The most basic form and structure of all life

 EXAMPLES OF CELLS

Amoeba Proteus

Plant Stem

Bacteria

Red Blood Cell

Nerve Cell
https://www.youtube.com/watch?v=URUJD5NEXC8
Animal and plant cell electron micrograph

Try to identify and label all organelles in both cells

LEARNING OBJECTIVES
• Outline key structural features of a prokaryotic cell as found in a typical
bacterium, including:
unicellular

generally 1–5 µm diameter

peptidoglycan cell walls

circular DNA

70S ribosomes

absence of organelles surrounded by double membranes.

 Compare the structure of a prokaryotic cell as found in a typical bacterium

with the structures of typical eukaryotic cells in plants and animals

https://www.youtube.com/watch?v=URUJD5NEXC8
Structure of a prokaryote cell
PROKARYOTES VS EUKARYOTES
Features Prokaryotes Eukaryotes

Size 1-5 µm in diameter Up to 40 µm in diameter

DNA Circular DNA lies free in DNA is contained in a

Naked DNA cytoplasm nucleus surrounded by
nuclear envelope
Ribosome Slightly smaller Slightly larger
(70s ribosomes – 20nm) (80s ribosomes – 25nm)
Compartmentalisation Very few organelles Many types of organelles
(non surrounded by double present
membrane)
[ Eg: lysosome & ribosome]
Cell wall Made of peptidoglycan Plants – cellulose/lignin
Fungi – chitin
Endoplasmic No ER present Has SER and RER
recticulum
 LEARNING OBJECTIVES
• Recognise organelles and other cell structures found in eukaryotic cells and
outline their structures and functions, limited to
cell surface membrane centrioles and microtubules
nucleus, nuclear envelope and nucleolus cilia
rough endoplasmic reticulum microvilli
smooth endoplasmic reticulum chloroplasts (including the presence of
small circular DNA)
Golgi body (Golgi apparatus or Golgi cell wall
complex)
mitochondria (including the presence of Cell wall
small circular DNA)
ribosomes (80S in the cytoplasm and 70S in plasmodesmata
chloroplasts and mitochondria).
lysosomes large permanent vacuole and tonoplast
of plant cells

• State that cells use ATP from respiration for energy-requiring processes
Overview of animal cell
 NUCLEUS (P: nuclei) • 5 -10 um
• the largest cell organelle
• Surrounded by double membrane
(nuclear envelope)
• Contain nuclear pores
• When the cell is not dividing,
chromosomes are loosely coiled
called chromatin
• Function: Contains the genetic
material
• responsible for protein synthesis,
cell division, controls cell activities

Nucleolus:
• Manufactures ribosomes and rRNA
• rRNA + protein => ribosome
Smooth ER
 ENDOPLASMIC RETICULUM
Rough endoplasmic reticulum Smooth endoplasmic reticulum
(RER) (SER)

Synthesize and transport of lipid

and steroids (cholesterol,
hormones)

 Transport protein Synthesize and transport of lipid and

 Has ribosomes steroids (cholesterol, hormones)
RIBOSOME

• Sites of protein synthesis

• Can be found free in cytoplasm or attached on the rough ER
• Very small: 25nm in diameter
• Made of ribonucleic acid (rRNA) and protein in the nucleolus
• Is found in mitochondria and chloroplast as well
• No membrane
Centrifugation

• S is a measurement unit svedbergs, used

to measure how fast molecules move in a
centrifuge.
 ENDOSYMBIONT THEORY
• “ENDO” – inside, “SYMBIONT” – organisms that lives in a
mutually beneficial relationship with another organism
• 1960s – scientist discovers that mitochondria and chloroplast
has ribosomes and small circular DNA
• Scientist proved that mitochondria and chloroplast are
ancient bacteria that now live in plants and animal cells
GOLGI BODY (aka: Golgi apparatus/Golgi complex)

Functions:

• Collects, processes and sorts

molecules (esp proteins from
RER)
• Transport molecules within or
out of the cell via Golgi vesicles
• Adding sugars to proteins
(glycosylation)
• Removing methionine (first
amino acid)
• Make lysosomes
• Convert sugars into cell wall
components (plants)
 LYSOSOME
• Single membrane
• Has no internal structure
• Contains digestive (hydrolytic)
enzyme

Functions: -
– breakdown material ingested
by phagocytic cells
– Breakdown unwanted
structures inside the cell or
the whole cell itself
IMPORTANCE OF LYSOSOME (EXAMPLE)
 MITOCHONDRION (p: mitochondria)

• Size: 1µm
• Double membrane
• Contains 70s ribosomes
• Has circular DNA in the matrix
• Function: Carry out aerobic respiration by producing ATP (adenosine
triphosphate). To synthesise lipid as well.
ATP (adenosine triphosphate): the molecule that is the universal energy currency in all
living cells; the purpose of respiration is to make ATP
CELL SURFACE MEMBRANE
• Size: 7nm
• Partially permeable
• Controls exchange between cell and its
environment

Trilaminar
appearance
CHLOROPLAST
• Size: 5µm
• Inside the Cytoplasm (Stroma), is
a system of layers of membranes
called grana. (s: granum)
• Grana made up of thylakoid
membranes, which contain light
absorbing pigments.
• Stroma is an organic matrix of
proteins, nucleic acids, DNA (small
circular) and enzymes.
• Following a period of illumination,
Photosynthesis takes place, and
Starch Grains & Lipid Globules
appear.
• Function: Photosynthesis takes
place in these organelles.
lamella
 MICROTUBULES
• Made of tubulin (protein) :- α- and β-
tubulin
• Tubulin dimers  protofilaments 
mictrotubules
• Long, rigid, hollow tubes in cytoplasm
• Determining cell shape (cytoskeleton)
and intracellular transport system
• Involved in nuclear division
• Form spindle fibres which attached to
centromere
• Spindle fibres shortening at anaphase
and moves sister chromatids to
opposite poles
• Size: 25 nm Ø https://www.youtube.com/watch?v=5rqbmLiSkpk
CENTROSOME
• The region where
centrioles align
• Believed to be the
MICROTUBULE
ORGANISING
CENTRES (MTOCs)
where the
microtubules
assembles
 CENTRIOLES

• Found in all animal cells in pair (2) (not in

plant cells)
• Hollow cylinders
• 9 sets of 3 microtubules
• Involved in nuclear division
• Size: 0.4 µm long
 CILIA (s. cilium)

SEM of cilia in the respiratory tract

Single celled organisms can therefore use the action of cilia

for locomotion
MICROVILLI (s. microvillus)

Increase surface area of the cell

surface membrane
CELL WALL
• Relatively rigid
• Contains fibres of
cellulose which
strengthens the wall
• Gives plant cell a
defined shape
• Prevents the plant cell
from bursting when
water enters by osmosis
• Fully permeable
• May be surrounded by
lignin
PLASMODESMATA (S: PLASMODESMA)

Linking the neighbouring

cells
 VACUOLES
Animals: phagocytic vacuoles
Plants: large, permanent, central vacuole

Surrounded by a membrane, called

tonoplast

Tonoplast: controls exchange between

cytoplasm and vacuole
Cell sap: contains mineral salts, sugars,
oxygen, CO2, pigments, enzymes and other
organic compounds and waste products
Function of vacuoles: help to regulate
osmotic properties of cells
LEARNING OUTCOMES
Use an eyepiece graticule and stage
micrometer scale to make and use
the appropriate units, millimetre
(mm), micrometre (µm) and
nanometre (nm)
 MEASURING CELLS

Eyepiece graticule

Stage micrometer
MEASUREMENT OF MICROSCOPE SAMPLES

A stage micrometer is a glass slide with etchings of

a known or measurable length.
Think of it as a tiny ruler
TRY THIS!!

mm μm nm
2 mm 2 2000 2000 000

130μm 0.13 130 130 000

0.032m 32 32 000 32 000 000

7.25μm 0.007 7.25 7250

 MICROMETRY
Steps:
1. Place the micrometer on the microscope stage
2. superimpose or align the graticule scale onto it.
Place your sample under the same magnification and line it
against the graticule
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0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8

Each division= 0.05 mm
mm

0 0.657 mm

100 divisions of the graticule= 0.657 mm

1 division
35 = 0.657mm
divisions= /100229.95 µm
35 x 6.57 µm=
=0.00657 mm or 6.57 µm
 Try this exercises

Smallest markings on SMS is 0.01mm apart

Calculate the actual width of a single cell.
Width of a single cell. (23 units)
(80) EG markings = (18) stage micrometer
Therefore, 1 unit of EGS = (18 x 0.01mm / 80 x 100μm = 2.25 μm)
Actual width= 23 units x 2.25 μm = 51.75 μm
 Do Now
The diagram shows a stage micrometer viewed with
an eyepiece graticule scale, using a magnification of
×400.

Using the same magnification, a chloroplast is

measured and found to be 8 eyepiece graticule
divisions long. How long is the chloroplast?
LEARNING OUTCOMES
State that all viruses are non-
cellular structures with a nucleic
acid core (either DNA or RNA) and a
capsid made of protein, and that
some viruses have an outer
envelope made of phospholipids
 Viruses “tiny non-cellular structures”

• Do not have cell structure (no plasma membrane)

• Simpler in structure
• Self-replicating molecule of DNA/RNA in host cell
• A protective protein coat (capsid) made up of capsomere
• Some virus have membranous envelope surrounding capsids derived from host
cell membrane (phospholipid).
• Can only reproduce by infecting and taking over living cells (parasitic)
• Size: 20-300nm
Coronavirus (RNA) Adenovirus (DNA)