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Comparison between TEM and SEM

Transmission Electron Microscopy and Scanning Electron Microscopy

10/18/2013 Amity school of Biotechnology Amit K umar Yadav, BTech.BioTech- 3rd semester

Comparison between TEM and SEM

Comparison includes similarities and dissimilarities. 1. Similarities between TEM and SEM A) The illumination source Both, TEM and SEM uses tungsten filament surrounded by a cathode shield as electron source or light. The light source or electron source is called as electron gun. The wavelength and voltage relation for TEM and SEM Wavelength=1.51/2/V nm 2. The lenses In both TEM and SEM, lenses are made of electromagnets. A variable electric current produces electric field of variable electric strength which focuses or defocuses electron beam on the specimen. 3. The vacuum system Vacuum system in TEM and SEM is primary requirement. The presence of air molecule results in the collision and scattering of electron from the path. Vacuum is maintained by high vacuum pumps.

Vacuum factor for TEM and SEM: biological materials must be properly fixed and preserved. 4. Preparation of biological materials of TEM and SEM observation In both TEM and SEM preparation is approximately same. Preparation includes fixation, dehydration, embedding, ultramicrotomy, staining and shadowing. 5. Other Similarities Apart from light microscopy, cell fractionation method etc. TEM and SEM are both a strong cell study technique. For both TEM and SEM Contrast is directly proportional to atomic number of molecule in the specimen. For both TEM and SEM, our sample must be conductive. IN both TEM and SEM, a high access voltage is required (100~400 mV).

B) Dissimilarities between TEM and SEM

1. Image Formation

Image formation in TEM is due to the scattering of electron while in SEM image is the result of secondary electron emission. In TEM scattering results in shadow formation on viewing screen or photographic film while in SEM, the secondary electron strike on scintillator and the light produced is converted to electric signals and after amplifying, it is displayed on Cathode Ray Tube (CRT). 2. Some Other Dissimllarities TEM gives internal structure while SEM is preferred for surface study. In TEM specimen is fixed (2D image) while in SEM, specimen is free to rotate (3D image). In TEM, there is no emission of any rays except electron beam while image formation. But in SEM, X-rays, gamma rays, beta rays can be emitted withn the secondary electron whiler image formation. For TEM horizontal resolution is 0.2nm and 5nm for SEM. Field of viewe for TEM is 100nm while for SEM it is 1mm.


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