2-Photon Imaging: From Polymeric Materials to Engineered Tissues

Lead Research Organisation: University of Sheffield
Department Name: Materials Science and Engineering

Abstract

Two-photon microscopy is a leading-edge imaging technology and a powerful research tool that combines long wavelength excitation and laser scanning microscopy. Of importance to our work it can enable capture of high resolution three dimensional images of living cells within 3D constructs as well as in-depth penetration of specimens tagged with very specific fluorophores. This technology is now becoming a method of choice for the dynamic imaging of biological and polymeric systems, not otherwise possible by other optical approaches and therefore will underpin a broad number of research programmes in biomaterials and tissue engineering.

Technical Summary

The current proposal is for the single purchase of a titanium-sapphire femtosecond pulsed NIR laser and a beam splitter to add to two existing confocal microscopes that are currently located in different departments. One system is inverted and equipped for fluorescence correlation spectroscopy (FCS) and the other is upright. Relocation of these will enable us to establish a single-site multiphoton / FCS imaging facility within the Kroto Research Institute at Sheffield University. This proposal outlines a number of highly developed and interconnected themes from 14 applicants that extend across the interface between the physical and life sciences, spanning polymeric biomaterials to tissue engineering, and justifies why a parallel two-photon confocal microscope facility with FCS is needed to develop our research programmes. This is most effectively answered by requesting money for a central multi-user facility that will have extensive multidisciplinary use and be based in a dedicated institute established for research across the disciplines.

Publications

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Murray-Dunning C (2008) Use of aligned polymer microfibres for peripheral nerve repair in 8th World Biomaterials Congress 2008

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Cantón I (2008) 3D tissue engineered constructs for real time detection of stress via NF-?B activation in transiently transfected human dermal fibroblasts in 8th World Biomaterials Congress 2008

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Filova E (2008) Mesenchymal stems cells and endothelial cells on micropatterned surfaces prepared by plasma polymerisation in 8th World Biomaterials Congress 2008

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Charnley M (2009) Generation of Bioactive Materials with Rapid Self-Assembling Resorcinarene-Peptides in Advanced Materials

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Baggaley E (2014) Dinuclear ruthenium(II) complexes as two-photon, time-resolved emission microscopy probes for cellular DNA. in Angewandte Chemie (International ed. in English)

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Bryan M (2010) Erratum: "The effect of trapping superparamagnetic beads on domain wall motion" [Appl. Phys. Lett. 96, 192503 (2010)] in Applied Physics Letters

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Chau DY (2013) The development of a 3D immunocompetent model of human skin. in Biofabrication

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Pateman CJ (2015) Nerve guides manufactured from photocurable polymers to aid peripheral nerve repair. in Biomaterials

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Hopper AP (2014) Amine functionalized nanodiamond promotes cellular adhesion, proliferation and neurite outgrowth. in Biomedical materials (Bristol, England)

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Sun T (2008) Development of a bioreactor for evaluating novel nerve conduits. in Biotechnology and bioengineering

 
Description The grant allowed the purchase of a versatile imaging system for conducting work on 3D in vitro cells/tissues. The major outcome has been the discovery of several methods for imaging a breadth of cells and scaffolds in 3D as a alternative to using animals.
Exploitation Route The grant has supported a tremendous breadth of outputs - not just those I am personally associated with (immediate outputs herein), but those of many other groups. These outputs have been documented on the webpage cited above and here - (https://www.sheffield.ac.uk/kroto/confocal).

These findings may be taken forward by devising new methods for further reducing animal research, new methods for SMEs to develop new implants, through to new bioreactors for such methods.
Sectors Healthcare
URL https://www.sheffield.ac.uk/kroto/confocal
 
Description This was an equipment grant for imaging 3D engineered tissues. It has led to a breadth of research represented in several outputs. Parallel findings have arisen including: 1. Design of commercial bioreactors in collaboration with SMEs 2. Design of methods for orthopaedic implants in collaboration with SMEs using 3D in vitro methods.
First Year Of Impact 2010
Sector Healthcare
Impact Types Economic