THE LIGHT MICROSCOPE AND THE

ELECTRON MICROSCOPE

Compound light microscope

© 2016 Paul Billiet ODWS Transmission electron microscope

 UNITS OF MEASUREMENT
1m = 103 mm (millimetres)
1m = 106 µm (micrometres)
1m = 109 nm (nanometres)

Sometimes in old texts Angstroms (Å) are used (the diameter of

a hydrogen atom)
1m = 1010 Å

© 2016 Paul Billiet ODWS

 The electromagnetic spectrum

© 2016 Paul Billiet ODWS

 Light vs Electron

© 2016 Paul Billiet ODWS

Magnifying
using the
Light
Microscope

© 2016 Paul Billiet ODWS

Magnifying
using the
Electron
Microscope

© 2016 Paul Billiet ODWS

 THE LIGHT MICROSCOPE v THE ELECTRON MICROSCOPE
FEATURE LIGHT MICROSCOPE ELECTRON MICROSCOPE

Electromagnetic Visible light Electrons

spectrum used 760nm (red) – 390nm app. 4nm
Colours visible Monochrome
Maximum app. 200nm 0.2nm
resolving power Fine detail
Maximum
magnification x1000 – x1500 x500 000
Radiation Tungsten or quartz High voltage (50kV)
source halogen lamp tungsten lamp
Lenses Glass Magnets
Interior Air-filled Vacuum
Focussing Human eye (retina), fluorescent (TV) screen,
screen photographic film photographic film
© 2016 Paul Billiet ODWS
THE LIGHT MICROSCOPE v THE ELECTRON MICROSCOPE

ELECTRON
FEATURE LIGHT MICROSCOPE
MICROSCOPE
Preparation of Temporary mounts Tissues must be
specimens living or dead dehydrated
= dead
Fixation Alcohol OsO4 or KMnO4
Embedding Wax Resin
Sectioning Hand or microtome Microtome only.
slices  20 000nm Slices  50nm
Whole cells visible Parts of cells visible
Stains Water soluble dyes Heavy metals

Support Glass slide Copper grid

© 2016 Paul Billiet ODWS
Microtome knife

© 2016 Paul Billiet ODWS © P Billiet

Copper grid slides

© 2016 Paul Billiet ODWS © P Billiet

 CELL STRUCTURE AND MICROSCOPY
SUBCELLULAR STRUCTURE EUKARYOTES PROKARYOTES
ANIMAL CELLS PLANT CELLS
Cell wall - but may have an + + but not of cellulose
extracellular matrix

Plasma membrane + cholesterol present + +

Nucleus & nucleoli + + -

Chromosomes + (DNA + Protein) + (DNA + Protein) Circular loop of DNA

Centrioles + - -

Mitochondria + + -

Plastids (chloroplasts, chromoplasts etc.) - + -

Sap vacuole - + -

Food vacuoles Some - -

Contractile vacuoles Some - -

Pseudopods Some - -
Cilia Some - -

Flagellae Some Some Rotating

Microtubules + + -

Lysosomes + - -

Golgi apparatus + + -

Microvilli Some - -
 CELL STRUCTURE AND MICROSCOPY

SUBCELLULAR EUKARYOTES PROKARYOTES

STRUCTURE
ANIMAL CELLS PLANT CELLS

Endoplasmic reticulum + + -
Nuclear envelope & pores + + -
Vesicles + + -
Ribosomes +(80S) +(80S) +(70S)
Organelle membranes + + -
Macromolecular structures + + +

© 2016 Paul Billiet ODWS