In which case does one use each of these techniques specifically for?
Hello Anahita,
Those two methods are quite different.
Rotary shadowing was successfully used for contrast enhancement in electron microscopy of nucleic acids (DNA, RNA, heteroduplexing, etc.). It can also be used for coating samples with conductive layer in scanning electron microscopy. For this method in principal, you need a special (ultra)high vacuum device with rotary stage and evaporator for heavy metals (platinum, platinum/carbon, paltinum:paladium alloy or platinum:iridium alloy).
There is an old book from eighties “Electron Microscopy in Molecular Biology: A Practical Approach (The Practical Approach Series) 1987 by J. Sommerville and U. Scheer (eds).” in which you can find a lot about the application of rotary shadowing.
The negative staining can be used for imaging of bacteriophages, bacteria, bacterial pili and flagella in TEM. I can be used for visualization of membrane vesicles, protein complexes, etc. It is really important method of diagnostic electron microscopy in virology (human, animal, plant). For negative staining you usually need uranyl acetate or uranyl formate or ammonium molybdate salt solutions or phosphotungstic acid solutions. An “UranyLess EM Stains” are also available.
There is a lot in the literature on negative staining, e.g.:
De Carlo, S. & Harris, J. R. (2011), 'Negative staining and cryo-negative staining of macromolecules and viruses for TEM.', Micron (Oxford, England : 1993) 42, 117- 131.
or
Hayat, M. A. (1986), Negative Staining 'Basic Techniques for Transmission Electron Microscopy', Elsevier BV, , pp. 232-264.
Regards
Oldrich
Thank you Oldřich Benada for that big chunk of information. I'm sure it will help me figure out the basic ideology of both these processes.
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts