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Integration of FISH and Microfluidics

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Book cover Fluorescence In-Situ Hybridization (FISH) for Microbial Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2246))

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Abstract

Suitable molecular methods for a faster microbial identification in food and clinical samples have been explored and optimized during the last decades. However, most molecular methods still rely on time-consuming enrichment steps prior to detection, so that the microbial load can be increased and reach the detection limit of the techniques.

In this chapter, we describe an integrated methodology that combines a microfluidic (lab-on-a-chip) platform, designed to concentrate cell suspensions and speed up the identification process in Saccharomyces cerevisiae , and a peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) protocol optimized and adapted to microfluidics. Microfluidic devices with different geometries were designed, based on computational fluid dynamics simulations, and subsequently fabricated in polydimethylsiloxane by soft lithography. The microfluidic designs and PNA-FISH procedure described here are easily adaptable for the detection of other microorganisms of similar size.

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Acknowledgments

This work was financially supported by: (1) Base Funding—UIDB/00511/2020 of the Laboratory for Process Engineering, Environment, Biotechnology and Energy—LEPABE—funded by national funds through the FCT/MCTES (PIDDAC); (2) Projects POCI-01-0145-FEDER-031011 (μFISH) and POCI-01-0145-FEDER-029961 (ColorISH), funded by FEDER funds through COMPETE2020—Programa Operacional Competitividade e Internacionalização (POCI) and by national funds (PIDDAC) through FCT/MCTES; and (3) BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020—Programa Operacional Regional do Norte.

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Authors and Affiliations

  1. LEPABE – Laboratory for Process Engineering, Environment, Biotechnology and Energy, Department of Chemical Engineering, Faculty of Engineering, University of Porto, Porto, Portugal

    Célia F. Rodrigues & Nuno F. Azevedo

  2. CEFT – Transport Phenomena Research Center, Department of Chemical Engineering, Faculty of Engineering, University of Porto, Porto, Portugal

    João M. Miranda

Authors
  1. Célia F. Rodrigues
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  2. Nuno F. Azevedo
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  3. João M. Miranda
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Corresponding author

Correspondence to João M. Miranda .

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Editors and Affiliations

  1. LEPABE – Laboratory for Process Engineering Environment, Biotechnology and Energy, Department of Chemical Engineering, University of Porto, Porto, Portugal

    Nuno F. Azevedo

  2. INIAV – National Institute for Agrarian and Veterinarian Research, Rua dos Lagidos, Lugar da Madalena, Vairão, CEB - Centre of Biological Engineering, University of Minho, Campus de Gualtar, Braga, LEPABE – Laboratory for Process Engineering Environment, Biotechnology and Energy, Department of Chemical Engineering, University of Porto, Vila do Conde, Portugal

    Carina Almeida

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Rodrigues, C.F., Azevedo, N.F., Miranda, J.M. (2021). Integration of FISH and Microfluidics. In: Azevedo, N.F., Almeida, C. (eds) Fluorescence In-Situ Hybridization (FISH) for Microbial Cells. Methods in Molecular Biology, vol 2246. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1115-9_16

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  • DOI: https://doi.org/10.1007/978-1-0716-1115-9_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1114-2

  • Online ISBN: 978-1-0716-1115-9

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