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Microscopic Detection of Yeasts Using Fluorescence In Situ Hybridization

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Fungal Diagnostics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 968))

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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Author information

Authors and Affiliations

  1. Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa (IHMT/UNL), Lisboa, Portugal

    João Inácio

  2. Instituto de Higiene e Medicina Tropical/Centro de Recursos Microbiológicos, Universidade Nova de Lisboa (IHMT/UNL), Lisboa, Portugal

    Maria da Luz Martins

Authors
  1. João Inácio
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  2. Maria da Luz Martins
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Corresponding author

Correspondence to Maria da Luz Martins .

Editor information

Editors and Affiliations

  1. , NCBES, Molecular Diagnostics Research Gr, National University of Ireland, Galway, IDA Business Park, Dangan, Galway, Ireland

    Louise O'Connor

  2. , NCBES, Molecular Diagnostics Research Gr, National University of Ireland, Galway, IDA Business Park, Dangan, Galway, Ireland

    Barry Glynn

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Inácio, J., da Luz Martins, M. (2013). Microscopic Detection of Yeasts Using Fluorescence In Situ Hybridization. In: O'Connor, L., Glynn, B. (eds) Fungal Diagnostics. Methods in Molecular Biology, vol 968. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-257-5_5

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  • DOI: https://doi.org/10.1007/978-1-62703-257-5_5

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-256-8

  • Online ISBN: 978-1-62703-257-5

  • eBook Packages: Springer Protocols

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