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Analysis of Extracellular Vesicles Using Fluorescence Nanoparticle Tracking Analysis

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Extracellular Vesicles

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1660))

Abstract

Fluorescence nanoparticle tracking analysis (fl-NTA) allows for accurate sizing, counting, and phenotyping of extracellular vesicles (EV). Here, we present two protocols for the analysis of EVs using fl-NTA, highlighting the potential pitfalls and challenges. The first protocol utilizes CellMask Orange™ (CMO) as a general membrane marker to label EVs derived from plasma. The second protocol describes the use of a Qdot-conjugated antibody to identify syncytiotrophoblast (STB)-derived EVs. “Standard” preparations of STB-derived EVs enriched for either microvesicles (STBMV) or exosomes (STBEX), containing a known amount of EV positive for the STB specific antigen placental alkaline phosphatase (PLAP), were also used to optimize fl-NTA camera settings.

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References

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Acknowledgment

This work was supported by MRC programme grant MR/J003360/1.

Author information

Authors and Affiliations

  1. Malvern Instruments Ltd, Amesbury, UK

    Pauline Carnell-Morris, Agnieszka Siupa & Patrick Hole

  2. Department of Food and Nutritional Sciences, University of Reading, Reading, RG6 6AP, UK

    Dionne Tannetta

  3. Nuffield Department of Obstetrics & Gynaecology, University of Oxford, Women’s Centre Level 3, John Radcliffe Hospital, Oxford, OX3 9DU, UK

    Rebecca Dragovic

Authors
  1. Pauline Carnell-Morris
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  2. Dionne Tannetta
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  3. Agnieszka Siupa
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  4. Patrick Hole
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  5. Rebecca Dragovic
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Corresponding author

Correspondence to Rebecca Dragovic .

Editor information

Editors and Affiliations

  1. CloudHealth Genomics, Ltd., Shanghai, China

    Winston Patrick Kuo

  2. Predicine, Inc., Hayward, California, USA

    Shidong Jia

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Carnell-Morris, P., Tannetta, D., Siupa, A., Hole, P., Dragovic, R. (2017). Analysis of Extracellular Vesicles Using Fluorescence Nanoparticle Tracking Analysis. In: Kuo, W., Jia, S. (eds) Extracellular Vesicles. Methods in Molecular Biology, vol 1660. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7253-1_13

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  • DOI: https://doi.org/10.1007/978-1-4939-7253-1_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7251-7

  • Online ISBN: 978-1-4939-7253-1

  • eBook Packages: Springer Protocols

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