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.
|Title of host publication||Fungal Diagnostics - Methods in Molecular Biology|
|Editors||L O'Connor, B Glynn|
|Place of Publication||Totowa, USA|
|Publication status||Published - 1 Jan 2013|