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

doi:10.1007/s00216-016-0036-5

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

Communities & Collections

pcg.skipToMenu

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

View Usage Statistics

Bibliographic description

punktacja MEiN [2017 A]: 40

title:	Immobilization and detection of platelet-derived extracellular vesicles on functionalized silicon substrate : cytometric and spectrometric approach
author:	Gajos Katarzyna , Kamińska Agnieszka , Awsiuk Kamil , Bajor Adrianna, Gruszczyński Krzysztof, Pawlak Anna , Żądło Andrzej , Kowalik Artur, Budkowski Andrzej , Stępień Ewa
journal title:	Analytical and Bioanalytical Chemistry
volume:	409
issue:	4
date of publication :	2017
pages:	1109-1119
ISSN:	1618-2642
eISSN:	1618-2650
DOI:	10.1007/s00216-016-0036-5
language:	English
journal language:	English
abstract in English:	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).
keywords in English:	time-of-flight secondary ion mass spectrometry, flow cytometry, biomarkers, extracellular vesicles, nanoparticles/nanotechnology
affiliation:	Wydział Fizyki, Astronomii i Informatyki Stosowanej : Instytut Fizyki im. Mariana Smoluchowskiego, Wydział Biochemii, Biofizyki i Biotechnologii : Zakład Biofizyki
type:	journal article
subtype:	academic paper