Four-wave mixing imaging to study protein entry and release in mammalian cells [Abstract]

Masia, Francesco ORCID: https://orcid.org/0000-0003-4958-410X, Langbein, Wolfgang Werner ORCID: https://orcid.org/0000-0001-9786-1023, Borri, Paola ORCID: https://orcid.org/0000-0002-7873-3314 and Watson, Peter Duncan ORCID: https://orcid.org/0000-0003-0250-7852 2010. Four-wave mixing imaging to study protein entry and release in mammalian cells [Abstract]. Drug Discovery Today 15 (23-24) , p. 1089. 10.1016/j.drudis.2010.09.377

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Abstract

Optical microscopy is a powerful tool for tracking the binding, internalisation and subcellular trafficking of delivery vectors to mammalian cells. By exploiting multiphoton processes, subcellular structures can be imaged with intrinsic three-dimensional (3D) spatial resolution. Common fluorescent labels in multiphoton microscopy include organic fluorophores, which suffer from photobleaching, and quantum dots which are more photostable but contain cytotoxic elements (such as Cd or In). Gold nanoparticles (GNPs) are ideal optical labels in terms of photostability and bio-compatibility, but emit weak fluorescent signal. We have developed a novel multiphoton microscopy technique that exploits the third-order nonlinearity called four-wave mixing (FWM) of GNPs in resonance with their surface plasmon. In terms of imaging performances, FWM microscopy features a spatial resolution better than the one-photon diffraction limit and optical sectioning capabilities. We show high-contrast background-free imaging of gold-labels (down to 5 nm size) and sensitivity to the single particle level. We are also able to demonstrate a directed dissociation of the GNP from bound proteins at their surface. These results pave the way for active tracking of conjugated nanoparticles, before the controlled release of therapeutically relevant proteins to a localised site of interest.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Physics and Astronomy Biosciences
Subjects:	Q Science > QC Physics Q Science > QH Natural history > QH301 Biology
Additional Information:	Delegate Abstracts: Drug Discovery Today, Volume 15, Issues 23–24, December 2010, Pages 1080-1114
Publisher:	Elsevier
ISSN:	1359-6446
Funders:	EPSRC
Last Modified:	06 Nov 2022 13:33
URI:	https://orca.cardiff.ac.uk/id/eprint/27332

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