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.
Organisations
Publications
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)
Botchway SW
(2008)
Time-resolved and two-photon emission imaging microscopy of live cells with inert platinum complexes.
in Proceedings of the National Academy of Sciences of the United States of America
Bryan M
(2010)
Erratum: "The effect of trapping superparamagnetic beads on domain wall motion" [Appl. Phys. Lett. 96, 192503 (2010)]
in Applied Physics Letters
Cantòn I
(2010)
Development of a 3D human in vitro skin co-culture model for detecting irritants in real-time.
in Biotechnology and bioengineering
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
Charnley M
(2009)
Generation of Bioactive Materials with Rapid Self-Assembling Resorcinarene-Peptides
in Advanced Materials
Chau DY
(2013)
The development of a 3D immunocompetent model of human skin.
in Biofabrication
Chunthapong J
(2008)
A 3D Skin tissue-engineered model for inflammatory and toxicity testing
in European Cells and Materials
Filova E
(2008)
Mesenchymal stems cells and endothelial cells on micropatterned surfaces prepared by plasma polymerisation
in 8th World Biomaterials Congress 2008
Hopper AP
(2014)
Amine functionalized nanodiamond promotes cellular adhesion, proliferation and neurite outgrowth.
in Biomedical materials (Bristol, England)
Hopper AP
(2016)
Photochemically modified diamond-like carbon surfaces for neural interfaces.
in Materials science & engineering. C, Materials for biological applications
Kaewkhaw R
(2009)
Adipose-derived stem cells (ASCs) for peripheral nerve repair
in European Cells and Materials
Lizarraga-Valderrama L
(2015)
Nerve tissue engineering using blends of poly(3-hydroxyalkanoates) for peripheral nerve regeneration
in Engineering in Life Sciences
Morris H
(2008)
The role of mechanotransduction in bone tissue engineering
in European Cells and Materials
Morris HL
(2010)
Mechanisms of fluid-flow-induced matrix production in bone tissue engineering.
in Proceedings of the Institution of Mechanical Engineers. Part H, Journal of engineering in medicine
Murray-Dunning C
(2008)
Peripheral nerve engineering using aligned polymer microfibres
in European Cells and Materials
Murray-Dunning C
(2008)
Use of aligned polymer microfibres for peripheral nerve repair
in 8th World Biomaterials Congress 2008
Murray-Dunning C
(2009)
The use of aligned polymer microfibres in peripheral nerve engineering
in European Cells and Materials
Pateman CJ
(2015)
Nerve guides manufactured from photocurable polymers to aid peripheral nerve repair.
in Biomaterials
Paviolo C
(2014)
Laser exposure of gold nanorods can induce intracellular calcium transients.
in Journal of biophotonics
Plenderleith RA
(2015)
Arginine-glycine-aspartic acid functional branched semi-interpenetrating hydrogels.
in Soft matter
| 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 |