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Flow Cytometry and Fluorescence Microscopy as Tools for Structural and Functional Analysis of Vacuoles Isolated from Yeast and Plant Cells

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Plant Vacuolar Trafficking

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1789))

Abstract

A series of optimized protocols to isolate vacuoles from both yeast and plant cells, and to characterize the purified organelles at a functional and structural level, are described. For this purpose, we took advantage of the combined use of cell fractionation techniques with different fluorescence-based approaches namely flow cytometry, fluorescence microscopy and spectrofluorimetry. These protocols altogether constitute valuable tools for the study of vacuole structure and function, as well as for the high-throughput screening of drug libraries to identify new molecules that target the vacuole.

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Author information

Author notes

  1. Jorge Rodrigues and Cátia Pereira contributed equally to this work.

Authors and Affiliations

  1. Centre of Molecular and Environmental Biology, CBMA, University of Minho, Braga, Portugal

    Jorge M. P. Rodrigues, Cátia S. Pereira, Hernâni Gerós & Manuela Côrte-Real

  2. Centre of Biological Engineering, CEB, University of Minho, Braga, Portugal

    Cátia S. Pereira & Hernâni Gerós

  3. Sogrape Vinhos, S.A., Research and Development Department, Aldeia Nova, Avintes, Portugal

    Natacha Fontes

  4. Centre for the Research and Technology of Agro-environmental and Biological Sciences, CITAB—UMinho Pole, University of Minho, Braga, Portugal

    Hernâni Gerós

Authors
  1. Jorge M. P. Rodrigues
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  2. Cátia S. Pereira
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  3. Natacha Fontes
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  4. Hernâni Gerós
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  5. Manuela Côrte-Real
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Corresponding author

Correspondence to Hernâni Gerós .

Editor information

Editors and Affiliations

  1. Faculty of Sciences, University of Porto, Porto, Portugal

    Cláudia Pereira

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Rodrigues, J.M.P., Pereira, C.S., Fontes, N., Gerós, H., Côrte-Real, M. (2018). Flow Cytometry and Fluorescence Microscopy as Tools for Structural and Functional Analysis of Vacuoles Isolated from Yeast and Plant Cells. In: Pereira, C. (eds) Plant Vacuolar Trafficking. Methods in Molecular Biology, vol 1789. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7856-4_8

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  • DOI: https://doi.org/10.1007/978-1-4939-7856-4_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7855-7

  • Online ISBN: 978-1-4939-7856-4

  • eBook Packages: Springer Protocols

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