4 Microscopy

Publication date: 2011 Source:Methods in Microbiology, Volume 38 Author(s): Manfred Rohde In this chapter, the emphasis is placed on light microscopic examinations of unstained and stained specimens, as well as preparation protocols for transmission electron microscopy (TEM) and field emission scanning electron microscopy (FESEM), with the intention to provide a concise hands-on guide for scientists who would gather information about the structure of microorganisms using light and electron microscopic techniques. First, illuminating modes in light microscopic examination of microorganisms are described, followed by preparation protocols for identification of microorganism components like flagella and staining of intracellular granules in bacteria. In addition, differential staining for identification of bacteria like Gram-staining or acid-fast staining is described. This is followed by two different methodologies for a fast and reproducible negative staining, a method used for detection of bacterial appendages like flagella, pili or fimbriae for electron microscopic examinations. In addition, metal-shadowing techniques are briefly described. For visualization of intracellular ultrastructure, methods for standard embedding procedures are given. Furthermore, a method is described to enhance the contrast of intracellular membranes. For immune cytochemical localization studies, the next part deals with the progressive lowering of temperature (PLT) embedding protocol for Lowi...

Source: Methods in Microbiology - October 12, 2014 Category: Microbiology Source Type: research