The light microscope

Cells and microscopy 1.1.d, 1.1.a, 1.1.c

Size and magnification calculations 1.1.b, 1.1.e
The Electron Microscope
A typical plant cell
A typical plant cell
A typical plant cell
 Resolution
• Ability of a microscope to distinguish two
objects as separate from one other
• The smaller & closer together the objects that
can be distinguished as separate, the higher
the resolution (resolving power)
• The resolving power of the light microscope is
limited by the wavelength of light. It cannot
resolve detail finer than 0.2µm
The importance of resolution

Light Microscope,
low resolution

Electron Microscope,
high resolution
 Chromosomes
Light
Microscope

Scanning Electron
Microscope
 Light Microscope: Anaphase

Animal Plant
Cell Cell
SEM (scanning electron microscope)
and TEM (transmission electron
microscope)
TEM – Spindle Fibres
SEM
Video
 Light vs. Electron microscopes
Feature Light microscope Electron microscope
Radiation used Light rays Electron beams
Magnification x 2000 x 500 000
Resolving power 200 nm 0.2nm
Focused by Glass lenses Electromagnets
Biological material Living or dead Dead
Size Small & portable Very large & static
Preparation of Quick & simple Time-consuming &
material complex
Cost Relatively cheap VERY expensive
 Measurements & equivalents
1 millimetre (mm) 10-3 metre (m) 1/1 000 m

1 micrometre (µm) 10-6 metre (m) 1/1 000 000 m

1 nanometre (nm) 10-9 metre (m) 1/ 1 000 000 000 m

 Measuring Cells
• To accurately measure cellular structures we
need a suitable scale
 Estimating cell size
Graticule = eypiece micrometer – a fine scale
that fits inside an eyepiece lens

Stage micrometer = slide with a fine scale of

known dimension etched onto it
 Comparing
relative sizes
of molecules
 Magnification
• The size of an image of an object compared to
its actual size.
• Calculated using the formula M = I/A

I = size of image I
A = actual size of object
M = magnification M A
BUT you must remember to
convert values to the same unit FIRST
 Calculations

M = I/A I=MxA
I
÷ ÷
MXA
A = I/M
 Eg. Calculating Magnification
It is essential that the same unit is used for the size
of the image and the size of the object:

• Eg. If an image measures 50mm (as printed on

paper) and the object actually measures 5µm;
The size of the image should be converted to µm:
Size of image = 50mm = 50 000µm
Then you can use this to calculate magnification:
Therefore, magnification = 50 000/5 = 10,000
 Practice Question:
Calculate the linear magnification
1. If a red blood cell has a diameter of 8 µm and
a student shows it with a diameter of 40 mm
in a drawing, what is the magnification of the
drawing?
 Another Practice Question;
2. An image of a liver cell has a real scale bar
next to it recording 10um, but you measure
the scale bar and find it is 20mm. What is the
magnification used?
ANSWER:
• 20mm = 20,000 um

• Magnification = 20,000 \ 10
= 2000 X