Fluorescence in situ Hybridization (FISH) for the Identification and Quantification of Microorganisms
Cristina Moraru and Elke Allers
from: Applications of Molecular Microbiological Methods (Edited by: Torben L. Skovhus, Sean M. Caffrey and Casey R.J. Hubert). Caister Academic Press, U.K. (2014)
Fluorescence in situ hybridization (FISH) targeting ribosomal RNA (rRNA) has become a standard method in molecular ecology. FISH allows identification and quantification of microorganisms. The main methodological variations are the use of fluorochrome labeled probes (monolabeled FISH, DOPE-FISH) or Horse Radish Peroxidase (HRP) labeled probes, known as Catalyzed Reporter Deposition-FISH (or CARD-FISH). The first section of this chapter provides an overview of (i) the two methodological variations, (ii) sample evaluation by microscopy and image analysis, and (iii) probe design and principles of specific hybridization. The second section discusses pros and cons of rRNA FISH, including a comparison of the monolabeled and CARD-FISH variations. The final section describes different applications and new developments in rRNA FISH, emphasizing the wide range of questions and samples it can be applied to read more ...