title: Microscopic detection of yeasts using fluorescence in situ hybridization.
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.
authors.affiliation: Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa (IHMT/UNL), Lisboa, Portugal.
authors.forename: João
authors.initials: J
authors.lastname: Inácio
authors.forename: Maria
authors.initials: M
authors.lastname: da Luz Martins
mesh_terms: D017088:AIDS-Related Opportunistic Infections / Q000134:cerebrospinal fluid; D002555:Cerebrospinal Fluid / Q000382:microbiology; D003455:Cryptococcus neoformans / Q000166:cytology / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification*; D046508:Culture Techniques; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*