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Enhanced fluorescence imaging of live cells by effective cytosolic delivery of probes

Massignani, M., Canton, I., Sun, T., Hearnden, V., MacNeil, S., Blanazs, A., Armes, S.P., Lewis, A. and Battaglia, G. (2010) Enhanced fluorescence imaging of live cells by effective cytosolic delivery of probes. Plos One, 5 (5). Art no.e10459. ISSN 1932-6203

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Abstract

Background: Microscopic techniques enable real-space imaging of complex biological events and processes. They have become an essential tool to confirm and complement hypotheses made by biomedical scientists and also allow the re-examination of existing models, hence influencing future investigations. Particularly imaging live cells is crucial for an improved understanding of dynamic biological processes, however hitherto live cell imaging has been limited by the necessity to introduce probes within a cell without altering its physiological and structural integrity. We demonstrate herein that this hurdle can be overcome by effective cytosolic delivery.

Principal Findings: We show the delivery within several types of mammalian cells using nanometre-sized biomimetic polymer vesicles (a.k.a. polymersomes) that offer both highly efficient cellular uptake and endolysomal escape capability without any effect on the cellular metabolic activity. Such biocompatible polymersomes can encapsulate various types of probes including cell membrane probes and nucleic acid probes as well as labelled nucleic acids, antibodies and quantum dots.

Significance: We show the delivery of sufficient quantities of probes to the cytosol, allowing sustained functional imaging of live cells over time periods of days to weeks. Finally the combination of such effective staining with three-dimensional imaging by confocal laser scanning microscopy allows cell imaging in complex three-dimensional environments under both mono-culture and co-culture conditions. Thus cell migration and proliferation can be studied in models that are much closer to the in vivo situation.

Item Type: Article
Copyright, Publisher and Additional Information: © 2010 Massignani et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Keywords: Polymer Vesticles; Gene-Expression; Living Cells; Microscopy; Skin; Copolymers; Nanoscopy; Progress
Institution: The University of Sheffield
Academic Units: The University of Sheffield > Faculty of Science (Sheffield) > School of Biological Sciences (Sheffield) > Department of Biomedical Science (Sheffield)
The University of Sheffield > Faculty of Engineering (Sheffield) > Department of Materials Science and Engineering (Sheffield)
The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > School of Dentistry (Sheffield)
The University of Sheffield > Faculty of Science (Sheffield) > Department of Chemistry (Sheffield)
Depositing User: Miss Anthea Tucker
Date Deposited: 24 May 2010 09:19
Last Modified: 27 Jun 2014 22:06
Published Version: http://dx.doi.org/10.1371/journal.pone.0010459
Status: Published
Publisher: Public Library Science
Refereed: Yes
Identification Number: 10.1371/journal.pone.0010459
URI: http://eprints.whiterose.ac.uk/id/eprint/10840

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