Labeling Extracellular Vesicles for Nanoscale Flow Cytometry

Scientific Reports
Aizea Morales-KastresanaJennifer C Jones

Abstract

Extracellular vesicles (EVs), including exosomes and microvesicles, are 30-800 nm vesicles that are released by most cell types, as biological packages for intercellular communication. Their importance in cancer and inflammation makes EVs and their cargo promising biomarkers of disease and cell-free therapeutic agents. Emerging high-resolution cytometric methods have created a pressing need for efficient fluorescent labeling procedures to visualize and detect EVs. Suitable labels must be bright enough for one EV to be detected without the generation of label-associated artifacts. To identify a strategy that robustly labels individual EVs, we used nanoFACS, a high-resolution flow cytometric method that utilizes light scattering and fluorescence parameters along with sample enumeration, to evaluate various labels. Specifically, we compared lipid-, protein-, and RNA-based staining methods and developed a robust EV staining strategy, with the amine-reactive fluorescent label, 5-(and-6)-Carboxyfluorescein Diacetate Succinimidyl Ester, and size exclusion chromatography to remove unconjugated label. By combining nanoFACS measurements of light scattering and fluorescence, we evaluated the sensitivity and specificity of EV labeling assa...Continue Reading

References

Feb 21, 2004·Methods in Molecular Biology·Stephen P PerfettoRichard A Koup
Jan 30, 2008·Current Protocols in Cell Biology·Clotilde ThéryAled Clayton
Jun 6, 2009·Nature Reviews. Immunology·Clotilde ThéryElodie Segura
Oct 26, 2011·Nanomedicine : Nanotechnology, Biology, and Medicine·Esther N M Nolte-'t HoenMarca H M Wauben
Jan 20, 2012·Journal of Visualized Experiments : JoVE·Cecilia LässerJan Lötvall
Apr 3, 2013·Current Protocols in Cytometry·A Bruce LyonsKathleen V Doherty
Sep 7, 2013·Journal of Extracellular Vesicles·Kenneth W WitwerFred Hochberg
Mar 13, 2014·Journal of Thrombosis and Haemostasis : JTH·N ArraudA R Brisson
Apr 23, 2014·Nature Biotechnology·Hyungsoon ImHakho Lee
Oct 4, 2014·Journal of Extracellular Vesicles·Anita N BöingRienk Nieuwland
Oct 8, 2014·Annual Review of Cell and Developmental Biology·Marina ColomboClotilde Théry
Feb 18, 2015·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Tom Groot KormelinkMarca H M Wauben
Apr 11, 2015·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Nicolas ArraudAlain R Brisson
Oct 21, 2015·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Samuel A StonerJohn P Nolan
Nov 3, 2015·Nature·Ayuko HoshinoDavid Lyden
Jan 26, 2016·ACS Nano·Sangmoo JeongHakho Lee
Mar 29, 2016·Journal of Extracellular Vesicles·Chang-Sook HongTheresa L Whiteside
Aug 16, 2016·Biomaterials·Dionysios C WatsonGeorge N Pavlakis
Dec 13, 2016·Methods in Molecular Biology·Aizea Morales-Kastresana, Jennifer C Jones

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Citations

Jul 13, 2017·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Shuangtao MaDonna H Wang
Dec 22, 2017·Nanoscale·Marcel I RamirezEmmanuel Dias-Neto
Apr 1, 2018·Immunology and Cell Biology·Mancy TongLawrence W Chamley
Jun 15, 2018·Chemical Communications : Chem Comm·Marco P MonopoliDonal F O'Shea
May 22, 2018·Journal of Thrombosis and Haemostasis : JTH·S F W M LibregtsM H M Wauben
Jul 17, 2018·Advanced Healthcare Materials·Wenshuo WangShutao Wang
Sep 17, 2018·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Masashi TakaoTetsuhiko Ohba
Nov 22, 2018·American Journal of Physiology. Cell Physiology·Annachiara MitrugnoOwen J T McCarty
Dec 19, 2018·Proteomics·Dongsic ChoiJanusz Rak
Feb 7, 2019·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Kelly McDanielFanyin Meng
Mar 21, 2019·Cellular and Molecular Life Sciences : CMLS·Marta Monguió-TortajadaFrancesc E Borràs
Apr 13, 2019·Annual Review of Analytical Chemistry·Hong LianXiaomei Yan
Aug 31, 2019·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Xinzhi LiColin D Funk
Oct 12, 2019·Proceedings of the National Academy of Sciences of the United States of America·Anand Kumar Rai, Patricia J Johnson
Jun 22, 2019·Annual Review of Biochemistry·D Michiel Pegtel, Stephen J Gould
Dec 13, 2019·Nature Reviews. Urology·Johannes Linxweiler, Kerstin Junker
Jan 23, 2020·International Journal of Molecular Sciences·Yong Weon YiByong Seung Cho
Oct 17, 2019·IUBMB Life·Soheila MohammadiHamed Mirzaei
Sep 13, 2019·Journal of Neuroimmune Pharmacology : the Official Journal of the Society on NeuroImmune Pharmacology·Ryan P McNamara, Dirk P Dittmer
May 7, 2020·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Conor C HorganMolly M Stevens
May 31, 2020·International Journal of Molecular Sciences·Irene C MarcuJasmin Bluteau
Jul 3, 2020·International Journal of Molecular Sciences·Francisca DiasRui Medeiros
Aug 1, 2018·Frontiers in Pharmacology·Mengliu Yang, Sherry Y Wu
Jan 17, 2019·Biochemical Society Transactions·Fabrice Lucien, Hon S Leong
Mar 29, 2019·Biological Procedures Online·Abir MondalAnjali Shiras
Jan 17, 2019·Journal of Biomedical Science·Chun-Yi Chiang, Chihchen Chen
Oct 22, 2019·European Journal of Immunology·Andrea CossarizzaArturo Zychlinsky
Jan 8, 2020·International Journal of Molecular Sciences·Fanny EnderFrank Gieseler
Jun 2, 2018·Clinical and Translational Medicine·R E LaneM Trau

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Methods Mentioned

BETA
flow cytometry
size exclusion chromatography
light scattering
Fluorescence
Confocal
flow

Software Mentioned

NTA
FlowJo
FlowJo TreeStar
GraphPad
nanoFACS
Zen
TreeStar

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