Effective Visualization and Easy Tracking of Extracellular Vesicles in Glioma Cells

Biological Procedures Online
Abir MondalAnjali Shiras

Abstract

Extracellular vesicles (EVs) are nano-sized, membrane-bound structures secreted by cells and play critical roles in mediating intercellular signaling. EVs based on their size as well as mechanisms of biosynthesis are categorized as either microvesicles (200-1000 nm) or exosomes (30-200 nm). The EVs carry several biomolecules like proteins, DNAs, RNAs, and lipids into other cells and modulate several cellular functions. Being of very small sizes, it is very challenging to analyze them using conventional microscopes. Here, we report a new method developed by us for visualizing EVs using simple immune-fluorescence based technique, wherein the isolated EVs can be stained with fluorescently tagged antibodies to proteins present in EVs. The stained EVs can then be analyzed by using either confocal or super-resolution microscopes. Our method detailed here is equally effective in staining proteins that are present inside the EVs as well as those localized to the membranes of vesicles. By employing unique staining strategies, we have minimized the background noise and thereby improved the signal strength in confocal microscope. Using electron microscopy, we have ascertained that the structural integrity of the labeled EVs is intact. Mor...Continue Reading

References

Nov 24, 2011·PloS One·Enrique ZudaireSonja Vermeren
Jun 30, 2012·Nature Methods·Johannes SchindelinAlbert Cardona
Feb 20, 2013·The Journal of Cell Biology·Graça Raposo, Willem Stoorvogel
Feb 11, 2014·Methods in Molecular Biology·Shuzhen GuoEng H Lo
Feb 28, 2014·International Journal of Molecular Medicine·Cristina GrangeGiovanni Camussi
Oct 8, 2014·Annual Review of Cell and Developmental Biology·Marina ColomboClotilde Théry
Feb 28, 2015·Seminars in Cell & Developmental Biology·Valentina R MinciacchiDolores Di Vizio
May 17, 2015·Journal of Extracellular Vesicles·María Yáñez-MóOlivier De Wever
Nov 3, 2015·Nature·Ayuko HoshinoDavid Lyden
Feb 24, 2016·Journal of Extracellular Vesicles·Nina KolihaStefan Wild
Apr 14, 2016·Scientific Reports·Mark A RiderDavid G Meckes
Jun 23, 2016·Journal of Extracellular Vesicles·Zoraida AndreuMaría Yáñez-Mó
Dec 14, 2016·Cancer Cell·Annette BeckerDavid Lyden
May 2, 2017·International Journal of Nanomedicine·Conary MeyerBiao Lu
May 13, 2017·Circulation Research·Frank A W CoumansRienk Nieuwland
May 14, 2017·Scientific Reports·Aizea Morales-KastresanaJennifer C Jones
Jul 15, 2017·Annals of Translational Medicine·Istvan FuriGyongyi Szabo
Aug 18, 2017·Journal of Extracellular Vesicles·Lucas TrepsJulie Gavard
Dec 1, 2017·Journal of Extracellular Vesicles·Kaloyan TakovSean M Davidson
Dec 30, 2017·ACS Nano·Kyungheon LeeRalph Weissleder
Mar 20, 2018·Biochimica Et Biophysica Acta. Biomembranes·Pia Pužar DominkušMetka Lenassi
Apr 18, 2018·BioMed Research International·Maria Yu KonoshenkoPavel P Laktionov

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Citations

Dec 7, 2019·Current Pharmaceutical Design·Katarzyna KluszczyńskaMarkus Düchler
May 28, 2020·Nature Reviews. Molecular Cell Biology·Killian O'BrienXandra O Breakefield
Oct 14, 2020·Tissue Engineering. Part a·Elham PishavarSang Jin Lee
Sep 30, 2020·Biochimica Et Biophysica Acta. General Subjects·Federico ColomboEmanuele Cocucci
Apr 4, 2021·Developmental Cell·Bong Hwan SungAlissa M Weaver
May 1, 2021·International Journal of Molecular Sciences·Allison MathiesenAnca Dobrian
Jun 3, 2021·International Journal of Molecular Sciences·Rabab N HamzahRobert J Griffin
Nov 23, 2021·ACS Biomaterials Science & Engineering·Johana UribeSusan Daniel

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Methods Mentioned

BETA
electron
atomic force microscopy
transfection
confocal microscopy
transmission electron microscopy
Assay
size-exclusion chromatography
flow cytometry

Software Mentioned

AngioTool64
dSTORM
AngioTool
ImageJ
Fiji
dSTORM Nanoimager
GraphPad Prism

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