|Burghardt, Robert - TX A&M UNIVERSITY|
Submitted to: Current Protocols in Microbiology
Publication Type: Book / Chapter
Publication Acceptance Date: May 20, 2006
Publication Date: November 20, 2006
Citation: Burghardt, R.C., Droleskey, R.E. 2006. Transmission electron microscopy. In: Coico, R., Kowalik, T., Quarles, J., Stevenson, B., Taylor, R., editors. Current Protocols in Microbiology. Hoboken, NJ: John Wiley and Sons, Inc. p. 2B.1.1-2B.1.39. Interpretive Summary: From the very start of the field of microbiology, microbiologists have used the light microscope to not only classify bacteria based on what they looked like but also to study how they interact with the cells they infect. With the invention of the transmission electron microscope in the 1930’s, microbiologists were not only been able to better see bacteria, but also viruses for the first time. Since that time the use of electron microscopes in the field of microbiology has become widespread. While they have the ability to see small viruses, bacteria and cells, these things must be prepared using special procedures in order to be viewed using an electron microscope. These procedures can vary depending on the source of the sample to be viewed along with the kind of information that is to be gathered about the sample. This chapter details procedures in a step by step manner for the preparation of a variety of samples that the average microbiologist might encounter for viewing in the transmission electron microscope.
Technical Abstract: Transmission electron microscopy (TEM) has long been an important tool in the expanding field of microbiology. A major virtue of TEM is the increased resolution afforded by the use of accelerated electron beams at wavelengths significantly shorter than visible light. When coupled with suitable preparative techniques, TEM allows researchers to identify and characterize microorganisms and the cells that they infect. These identifications and characterizations are typically performed using techniques based on negative staining, immuno-gold negative staining and thin sectioning of embedded cells and tissue. Preparation of microbiological based material for evaluation using these techniques frequently requires the adaptation of existing preparative procedures for other types of samples that have been previously reported in the literature. This chapter details useful and practical protocols in the preparation of microbiological samples for evaluation in the transmission electron microscope providing all information needed for their successful observation.