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Dynamic colocalization microscopy to characterize intracellular trafficking of nanomedicines

Vercauteren, Dries, Deschout, Hendrik, Remaut, Katrien, Engbersen, Johan F.J., Jones, Arwyn Tomos, Demeester, Jo, De Smedt, Stefaan C. and Braeckmans, Kevin 2011. Dynamic colocalization microscopy to characterize intracellular trafficking of nanomedicines. ACS Nano 5 (10) , pp. 7874-7884. 10.1021/nn2020858

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Abstract

To gain a better understanding of intracellular processing of nanomedicines, we employed quantitative live-cell fluorescence colocalization microscopy to study endosomal trafficking of polyplexes in retinal pigment epithelium cells. A new, dynamic colocalization algorithm was developed, based on particle tracking and trajectory correlation, allowing for spatiotemporal characterization of internalized polyplexes in comparison with endosomal compartments labeled with EGFP constructs. This revealed early trafficking of the polyplexes specifically to Rab5- and flotillin-2-positive vesicles and subsequent delivery to Rab7 and LAMP1-labeled late endolysosomes where the major fraction of the polyplexes remains entrapped for days, suggesting the functional loss of these nanomedicines. Colocalization of polyplexes with the autophagy marker LC3 suggests for the first time that the process of xenophagy could play an important role in the persistent endosomal entrapment of nanomedicines.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Pharmacy
Subjects: R Medicine > RS Pharmacy and materia medica
Uncontrolled Keywords: nanomedicines; nonviral ocular gene therapy; particle tracking; intracellular trafficking; colocalization; autophagy
Publisher: ACS Publications
ISSN: 1936-0851
Last Modified: 04 Jun 2017 03:41
URI: http://orca-mwe.cf.ac.uk/id/eprint/24870

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