Physical interpretation of the size and concentration of - - PowerPoint PPT Presentation

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Physical interpretation of the size and concentration of - - PowerPoint PPT Presentation

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Physical interpretation of the size and concentration of extracellular vesicles measured by advanced techniques Edwin van der Pol 1 Chris Gardiner 2 , Paul Harrison 3 , Guus Sturk 1 , T on van Leeuwen 1 , and Rienk Nieuwland 1 July 2nd, 2013 1

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Physical interpretation of the size and concentration of extracellular vesicles measured by advanced techniques

Edwin van der Pol1

1 Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands; 2 John Radcliffe Hospital, Oxford, UK; 3 University of Birmingham Medical School, Birmingham, UK;

July 2nd, 2013 Chris Gardiner2, Paul Harrison3, Guus Sturk1, T

  • n van Leeuwen1,

and Rienk Nieuwland1

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SLIDE 2

Presentation includes discussion of the following off-label use of a drug or medical device: N/A Disclosures of: Edwin van der Pol

Employment No conflict of interest to disclose Research support No conflict of interest to disclose Scientific advisory board No conflict of interest to disclose Consultancy No conflict of interest to disclose Speakers bureau No conflict of interest to disclose M ajor stockholder No conflict of interest to disclose Patents No conflict of interest to disclose Honoraria No conflict of interest to disclose Travel support No conflict of interest to disclose Other No conflict of interest to disclose

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Introduction

vesicles measured by different techniques* each technique obtains a different size distribution

* van der Pol et al. P-M O-405 (ISTH 2011)

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enable data comparison between techniques

  • btain the size distribution of vesicles

Goals

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standard population of

polystyrene beads urine vesicles

analyzed by

transmission electron microscopy

  • ld flow cytometer (FACSCalibur)

new flow cytometer (Apogee A50-Micro) nanoparticle tracking analysis (Nanosight NS500) resistive pulse sensing (Izon qNano)

M ethods

van der Pol et al. JTH 2010

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Transmission electron microscopy

500 nm

vesicle standard reference beads >10x >1,000x earth moon >10,000x

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Conventional flow cytometry

vesicle standard reference beads refractive index polystyrene = 1.61 refractive index vesicles* = 1.4

* van der Pol SC07-4.01 (ISTH 2013)

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Novel flow cytometry

vesicle standard reference beads refractive index polystyrene = 1.61 refractive index vesicles* = 1.4

* van der Pol SC07-4.01 (ISTH 2013)

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Nanoparticle tracking analysis

vesicle standard reference beads

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Resistive pulse sensing

vesicle standard reference beads

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Summary

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Summary

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Outlook – data comparison

transmission electron microscopy resistive pulse sensing

12% 80%

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Outlook – fit parameters for clinical studies

transmission electron microscopy

m

d d k d C

−

        = ) (

plankton power-law function:

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Academic M edical Center

Anita Böing Anita Grootemaat Chi Hau Frank Coumans M arianne Schaap Najat Hajji René Berckmans Henk van Veen Y uana Y uana

Acknowledgements

More on microparticle detection: edwinvanderpol.com Apogee flow systems Ltd.

Oliver Kenyon

Izon Ltd.

Hans van der Voorn Rebecca Warr

NanoSight Ltd.

Jonathan Smith Andrew Malloy