Electron Microscope. Light vs EM Which is which? What are the main differences?

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Presentation transcript:

Electron Microscope

Light vs EM Which is which? What are the main differences?

SEM vs TEM A seed viewed with a scanning EM

Learning Objectives Know the difference between a transmission and a scanning electron microscope Explain the benefits of electron microscopes Compare and contrast EM & LM

Electron Microscope EM generates beam of electrons Electron beam passes through very thin prepared sample Resolution is 0.5nm

Preparing Slide of EM Fix specimen in gluteraldehyde Dehydrate with ethanol Embed in resin Slice thinly Stain (salts of heavy metals – eg lead, uranium) Mount on copper grid Place in vacuum (allows electrons to travel towards specimen)

Transmission Electron Microscope Electrons pass through denser part of sample less easily so create contrast 2D Magnification up to x

Scanning Electron Microscope Electrons ‘bounce’ off metal-salt-stained sample 3D Magnification x

Electron MicroscopesLight Microscopes Maximum resolution is 0.5nm Maximum resolution is 200nm Useful magnification is up to 250,000x in TEM, 100,000x in SEM Useful magnification is around 1000x (1500x at best) Wavelength is 1.0nm.Wavelength is between nm. Highly detailed images, and even 3D surface imaging. See reasonable detail, with true colours. Can see organelles of cells, bacteria and even viruses. Good for small organisms, invertebrates and whole cells. Comparision of EM and LM

Advantages/Disadvantages of EM Advantages of EMDisadvantages of EM Resolution is x2000 more than LM Samples have to be placed in a vacuum Produces detailed images Very expensive SEM produces 3D images Need to be highly skilled to create samples