Electron Microscopy ! • Much better at magnifying than light microscopes. • Can magnify up to 2 million times. • Very expensive • Need special training to use.
Artefacts: g A visible structural detail caused by Electrons have a much processing the specimen smaller wavelength than and not by actually light waves ! featuring !
Transmission electron microscope !
A beam of electrons is transmitted through a specimen
and focussed to produce an image ↳ Best resolution power (0.5 nm)!
Only thin Results in denser parts of a
specimens! specimen appearing darker, as they absorb more electrons ! Scanning Electron Microscope ! A beam of electrons is sent across the surface of the specimen and reflected electrons are collected in a cathode ray tube to form an image. ↳ Has a greater field of depth so can provide 3D images!
Laser scanning confocal Lower resolution:
microscopy I 3-10 nm
This microscope moves a
single spot of focussed light across a specimen (point of illumination). This causes fluorescence from the components labelled with a ‘dye’. - The emitted light is filtered through a pin hole and only light that is very close to the focal plane is detected! L
Light emitted from other parts of the specimen would
reduce resolution and cause blurring. This radiation does not pass through and is therefore not detected! Uses now and in the future!
Now - diagnosis of eye diseases, drug development,
Fluorescent Tags ! ↳ By utilising antibodies with ‘tags’, special features can be targeted and studied with much more precision than with a staining technique. Summary Questions !
1. Electron microscopes have a much higher
resolution due to the beams of electrons having shorter wavelengths than those of a light microscope.~ 2.Artefacts are visible structures caused by processing of a specimen, like staining techniques. ~ 3.Much more complex sample prep methods which causes more damage to the samples, which results in the artefacts. ~ 4.Transmission electron microscope - 2D ~ 5/5 5.Scanning emission microscope - 3D ~ Laser scanning confocal microscopy
IOW high high high
colour no-colour Colour colour 2000 X 2 Mill X 500 000 X 17000 X ,
zoomm 0 5nM .
3-10 Am 200 M Short medium medium long
good bad bac bad
Visible A beam of Beam of This microscope moves a single light is electrons is electrons sent spot of focussed passed transmitted across surface light across a through through a of the specimen, specimen. This causes the specimen reflected fluorescence from specimen and electrons components with ‘dye’. and bent focussed to collected in The emitted light through a produce an cathode ray is filtered through a pin hole, only lens image! tube to form an light that is very system! image! close to the focal plane is detected!
