Unit 1.

Microscopy in cell studies

Magnification: number of times larger an image is compared to the real size of the object.
• Depends on the power of the objective and eyepiece lens used.
Resolution: ability to distinguish between two separate points. The amount of detail that can
be seen- higher resolution, higher detail.
Limit of resolution: half the wavelength of radiation used to view specimen.
• Electrons have extremely short wavelength.
• They’re negatively charged, thus easily focused using electromagnets.

Types of microscopes:

Feature Light Electron

Source of radiation Light Electrons

±0.005nm
Wavelength of radiation 400-700nm

Max resolution 200nm 0.0-0.5

Lens Glass Electromagnet

Specimen Alive Dead

Stains Coloured dyes Heavy metal

Black and white

Image Coloured photomicrograph
electronmicrograph

View Eye piece Fluorescent screen

- Vacuum present in EM to prevent electrons from colliding with air particles to gain a sharp image.
- Water boils in RT in a vacuum, so specimen should be dead.

Calculating magni ication:

Actual size =
Image size
Magni ication
f

Calculation example

0.1mm of SM = 40 div of EG

1 div of EG= 0.1/40

0.0025mm*1000= 2.5µm

2.5µm*4 egu of chloroplast width= 10µm

 Unit 1.2

Basic unit of living organisms

In an electron micrograph:

• Very small particles can be observed as the electrons are easily absorbed.
• The parts of the specimen that appear darker in the final image are denser and absorb more
electrons.
• Due to higher resolution, the electron micrographs of plant and animal cells show most
organelles
• Ultrastructure: the structure revealed by the electron micrograph.

• Organelles: functionally and structurally distinct part of a cell, usually membrane bound.

A generalised animal cell (20µm):

Cell surface membrane (7nm): a selectively permeable membrane in plant and animal cells that allows for the exchange of
certain biological molecules and ions.
o Extremely thin with trilaminar appearance
o It is comprised of phospholipid bilayers which are
assembled with the hydrophilic phosphate heads facing the aqueous environment (inside and outside the cell) and the
hydrophobic tails facing each other.
o Function:
▪Barrier between cytoplasm and external environment ▪Cell signalling
▪Cell recognition (surface antigens)
▪Cell-to-cell adhesion
▪Site for enzyme catalysed reactions
▪Anchoring the cytoskeleton
▪Selection of substances that enter/leave the cell ▪Formation of Hydrogen bonds with water for stability