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Immobilization and detection of platelet-derived extracellular vesicles on functionalized silicon substrate : cytometric and spectrometric approach

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Immobilization and detection of platelet-derived extracellular vesicles on functionalized silicon substrate : cytometric and spectrometric approach

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dc.contributor.author Gajos, Katarzyna [SAP14012070] pl
dc.contributor.author Kamińska, Agnieszka [SAP14013162] pl
dc.contributor.author Awsiuk, Kamil [SAP14001148] pl
dc.contributor.author Bajor, Adrianna pl
dc.contributor.author Gruszczyński, Krzysztof pl
dc.contributor.author Pawlak, Anna [SAP11016784] pl
dc.contributor.author Żądło, Andrzej [SAP13902010] pl
dc.contributor.author Kowalik, Artur pl
dc.contributor.author Budkowski, Andrzej [SAP11011875] pl
dc.contributor.author Stępień, Ewa [SAP14015168] pl
dc.date.accessioned 2017-02-28T09:28:42Z
dc.date.available 2017-02-28T09:28:42Z
dc.date.issued 2017 pl
dc.identifier.issn 1618-2642 pl
dc.identifier.uri http://ruj.uj.edu.pl/xmlui/handle/item/38951
dc.language eng pl
dc.rights Udzielam licencji. Uznanie autorstwa 4.0 Międzynarodowa *
dc.rights.uri http://creativecommons.org/licenses/by/4.0/pl/legalcode *
dc.title Immobilization and detection of platelet-derived extracellular vesicles on functionalized silicon substrate : cytometric and spectrometric approach pl
dc.type JournalArticle pl
dc.description.physical 1109-1119 pl
dc.abstract.en Among the various biomarkers that are used to diagnose or monitor disease, extracellular vesicles (EVs) represent one of the most promising targets in the development of new therapeutic strategies and the application of new diagnostic methods. The detection of circulating platelet-derived microvesicles (PMVs) is a considerable challenge for laboratory diagnostics, especially in the preliminary phase of a disease. In this study, we present a multistep approach to immobilizing and detecting PMVs in biological samples (microvesicles generated from activated platelets and human platelet-poor plasma) on functionalized silicon substrate. We describe the application of time-of-flight secondary ion mass spectrometry (TOF-SIMS) and spectroscopic ellipsometry methods to the detection of immobilized PMVs in the context of a novel imaging flow cytometry (ISX) technique and atomic force microscopy (AFM). This novel approach allowed us to confirm the presence of the abundant microvesicle phospholipids phosphatidylserine (PS) and phosphatidylethanolamine (PE) on a surface with immobilized PMVs. Phosphatidylcholine groups (C_{5}H_{12}N^{+}; C_{5}H_{15}PNO{_{4}}^{+}) were also detected. Moreover, we were able to show that ellipsometry permitted the immobilization of PMVs on a functionalized surface to be evaluated. The sensitivity of the ISX technique depends on the size and refractive index of the analyzed microvesicles.Graphical abstractHuman platelets activated with thrombin (in concentration 1IU/mL) generate population of PMVs (platelet derived microvesicles), which can be detected and enumerated with fluorescent-label method (imaging cytometry). Alternatively, PMVs can be immobilized on the modified silicon substrate which is functionalized with a specific IgM murine monoclonal antibody against human glycoprotein IIb/IIIa complex (PAC-1). Immobilized PMVs can be subjected to label-free analyses by means ellipsometry, atomic force microscopy (AFM) and time-of-flight secondary ion mass spectrometry (TOF-SIMS). pl
dc.subject.en time-of-flight secondary ion mass spectrometry pl
dc.subject.en flow cytometry pl
dc.subject.en biomarkers pl
dc.subject.en extracellular vesicles pl
dc.subject.en nanoparticles/nanotechnology pl
dc.description.volume 409 pl
dc.description.number 4 pl
dc.identifier.doi 10.1007/s00216-016-0036-5 pl
dc.identifier.eissn 1618-2650 pl
dc.title.journal Analytical and Bioanalytical Chemistry pl
dc.language.container eng pl
dc.affiliation Wydział Fizyki, Astronomii i Informatyki Stosowanej : Instytut Fizyki im. Mariana Smoluchowskiego pl
dc.affiliation Wydział Biochemii, Biofizyki i Biotechnologii : Zakład Biofizyki pl
dc.subtype Article pl
dc.rights.original CC-BY; inne; ostateczna wersja wydawcy; w momencie opublikowania; 0; pl
dc.identifier.project ROD UJ / P pl
.pointsMNiSW [2017 A]: 40


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Udzielam licencji. Uznanie autorstwa 4.0 Międzynarodowa Except where otherwise noted, this item's license is described as Udzielam licencji. Uznanie autorstwa 4.0 Międzynarodowa