Chapter 2 : Structure of cells and organelles

2.1 Light and Electron Microscopes

Light & Electron Microscopes Microscope is used to produce a magnified image of an object or specimen. Important tool in the study of cell structure

Light & Electron Microscopes

Light microscope
(a) Light passes through a specimen (the object) & then through two sets of lenses, the objective lens & the eyepiece lens. (b) The lenses refract (bend) light to give a magnified image of the object. The image may be projected into the viewers eye or onto a photographic film, which is called photomicrograph / light micrograph. (c) The resolving power of the light microscope is limited by the wavelength of light. Its resolution is 200nm / 0.2 m. Only magnify objects up to about 1500 times without losing clarity.

Eyepiece Body Tube Revolving Nosepiece Objective Lens Stage Clips Diaphragm

Arm

Stage
Coarse Focus Fine Focus Base

Light

Electron microscope
(a) Uses a beam of electrons. (b) Is used for the detailed study of cells. (c) Electron beams have short wavelength. Hence, has greater resolving power & produce much higher effective magnification. (d) The resolution is 0.5 - 1 nm. (e) Two main types of electron microscope : i) transmission electron microscope (TEM) ii) scanning electron microscope (SEM)

Transmission Electron Microscope

Scanning Electron Microscope

Transmission electron microscope (TEM)

(a) is used to study the details of ultrastructure of cells. (b) Electron beams generated by a hot cathode is accelerated towards the specimen. (c) Electron beam passes through ultra-thin dehydrated sections of dead specimen. (d) It is placed in a vacuum to minimise electron scattering due to collision between electron and molecule in the air. (e) The sections are treated with heavy metals such as uranyl, lead acetate or osmium tetroxide. Electrons are absorbed by the heavily stained parts but pass through the lightly stained parts. This provides contrast between different parts of the specimen. (f) Electromagnets are used to bend & focus the electrons producing an image on fluorescent screen or photographic film. A two dimensional view of the specimen is produced. An electron micrograph is a photograph taken.

TEM Sample support in grid

Image of bacteria (Bacillus subtilis)

Scanning electron microscope (SEM)

(a) is used to produce a three-dimensional view of objects. (b) It is uses an electron beam to scan along the surface of a dead specimen which has been coated with a thin layer of heavy metal, for example, gold. (c) A large portion of the electrons are reflected. Some of the electrons absorbed excite other electrons at the surface to give off secondary electrons. (d) A recording of the emission from the specimen provides a picture of the specimen.

Image of an ant in SEM

Measurement
1 kilometer (km) = 1000 (103) metres 1 metre (m) = 100 (102) centimetres 1 metre (m) = 1000 (103) milimetres 1 millimetre (mm) = 1000 (103) micrometres (m / microns) 1 micrometre (m) = 1000 (103) nanometres (nm)

Magnification
Magnification is the number of times that an image is larger than the specimen and is usually given by the formula: magnification = size of image size of specimen