Cargando…

Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis

Cellular microvesicles and nanovesicles (exosomes) are involved in many disease processes and have major potential as biomarkers. However, developments in this area are constrained by limitations in the technology available for their measurement. Here we report on the use of fluorescence nanoparticl...

Descripción completa

Detalles Bibliográficos
Autores principales: Dragovic, Rebecca A., Gardiner, Christopher, Brooks, Alexandra S., Tannetta, Dionne S., Ferguson, David J.P., Hole, Patrick, Carr, Bob, Redman, Christopher W.G., Harris, Adrian L., Dobson, Peter J., Harrison, Paul, Sargent, Ian L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3280380/
https://www.ncbi.nlm.nih.gov/pubmed/21601655
http://dx.doi.org/10.1016/j.nano.2011.04.003
_version_ 1782223822023294976
author Dragovic, Rebecca A.
Gardiner, Christopher
Brooks, Alexandra S.
Tannetta, Dionne S.
Ferguson, David J.P.
Hole, Patrick
Carr, Bob
Redman, Christopher W.G.
Harris, Adrian L.
Dobson, Peter J.
Harrison, Paul
Sargent, Ian L.
author_facet Dragovic, Rebecca A.
Gardiner, Christopher
Brooks, Alexandra S.
Tannetta, Dionne S.
Ferguson, David J.P.
Hole, Patrick
Carr, Bob
Redman, Christopher W.G.
Harris, Adrian L.
Dobson, Peter J.
Harrison, Paul
Sargent, Ian L.
author_sort Dragovic, Rebecca A.
collection PubMed
description Cellular microvesicles and nanovesicles (exosomes) are involved in many disease processes and have major potential as biomarkers. However, developments in this area are constrained by limitations in the technology available for their measurement. Here we report on the use of fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles. In this system vesicles are visualized by light scattering using a light microscope. A video is taken, and the NTA software tracks the brownian motion of individual vesicles and calculates their size and total concentration. Using human placental vesicles and plasma, we have demonstrated that NTA can measure cellular vesicles as small as ∼50 nm and is far more sensitive than conventional flow cytometry (lower limit ∼300 nm). By combining NTA with fluorescence measurement we have demonstrated that vesicles can be labeled with specific antibody-conjugated quantum dots, allowing their phenotype to be determined. FROM THE CLINICAL EDITOR: The authors of this study utilized fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles, demonstrating that NTA is far more sensitive than conventional flow cytometry.
format Online
Article
Text
id pubmed-3280380
institution National Center for Biotechnology Information
language English
publishDate 2011
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-32803802012-02-24 Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis Dragovic, Rebecca A. Gardiner, Christopher Brooks, Alexandra S. Tannetta, Dionne S. Ferguson, David J.P. Hole, Patrick Carr, Bob Redman, Christopher W.G. Harris, Adrian L. Dobson, Peter J. Harrison, Paul Sargent, Ian L. Nanomedicine Research Article Cellular microvesicles and nanovesicles (exosomes) are involved in many disease processes and have major potential as biomarkers. However, developments in this area are constrained by limitations in the technology available for their measurement. Here we report on the use of fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles. In this system vesicles are visualized by light scattering using a light microscope. A video is taken, and the NTA software tracks the brownian motion of individual vesicles and calculates their size and total concentration. Using human placental vesicles and plasma, we have demonstrated that NTA can measure cellular vesicles as small as ∼50 nm and is far more sensitive than conventional flow cytometry (lower limit ∼300 nm). By combining NTA with fluorescence measurement we have demonstrated that vesicles can be labeled with specific antibody-conjugated quantum dots, allowing their phenotype to be determined. FROM THE CLINICAL EDITOR: The authors of this study utilized fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles, demonstrating that NTA is far more sensitive than conventional flow cytometry. Elsevier 2011-12 /pmc/articles/PMC3280380/ /pubmed/21601655 http://dx.doi.org/10.1016/j.nano.2011.04.003 Text en © 2011 Elsevier Inc. https://creativecommons.org/licenses/by/3.0/ Open Access under CC BY 3.0 (https://creativecommons.org/licenses/by/3.0/) license
spellingShingle Research Article
Dragovic, Rebecca A.
Gardiner, Christopher
Brooks, Alexandra S.
Tannetta, Dionne S.
Ferguson, David J.P.
Hole, Patrick
Carr, Bob
Redman, Christopher W.G.
Harris, Adrian L.
Dobson, Peter J.
Harrison, Paul
Sargent, Ian L.
Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis
title Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis
title_full Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis
title_fullStr Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis
title_full_unstemmed Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis
title_short Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis
title_sort sizing and phenotyping of cellular vesicles using nanoparticle tracking analysis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3280380/
https://www.ncbi.nlm.nih.gov/pubmed/21601655
http://dx.doi.org/10.1016/j.nano.2011.04.003
work_keys_str_mv AT dragovicrebeccaa sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis
AT gardinerchristopher sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis
AT brooksalexandras sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis
AT tannettadionnes sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis
AT fergusondavidjp sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis
AT holepatrick sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis
AT carrbob sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis
AT redmanchristopherwg sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis
AT harrisadrianl sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis
AT dobsonpeterj sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis
AT harrisonpaul sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis
AT sargentianl sizingandphenotypingofcellularvesiclesusingnanoparticletrackinganalysis