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        Microscopic Detection of Yeasts Using Fluorescence In Situ Hybridization

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        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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