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Detection of Viral RNA by Fluorescence in situ Hybridization (FISH)
Published on: May 5, 2012
Microscopic detection of yeasts using fluorescence in situ hybridization
João Inácio1, Maria da Luz Martins
1Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa (IHMT/UNL), Lisboa, Portugal.
Abstract:
Fluorescence in situ hybridization (FISH) has been widely used for the detection and identification of microorganisms in their natural environments. In this chapter we describe the use of a simple FISH-based protocol to detect and identify clinically relevant yeast species in culture and biological samples using Cryptococcus neoformans as a model. After fixation of cells with paraformaldehyde, the same are embedded in hybridization buffer containing specific fluorochrome-labeled oligonucleotide probes. After incubation and a subsequent washing step for removing unbound probes, samples are analyzed by epifluorescence microscopy.
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