| EPSRC Reference: |
EP/H027815/1 |
| Title: |
Ultra-sensitive spectroscopies and micromanipulation techniques for the study of biological processes at single cell and single molecule level |
| Principal Investigator: |
De Luca, Dr A |
| Other Investigators: |
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| Researcher Co-Investigators: |
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| Project Partners: |
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| Department: |
Physics and Astronomy |
| Organisation: |
University of St Andrews |
| Scheme: |
Postdoc Research Fellowship |
| Starts: |
01 October 2010 |
Ends: |
29 February 2012 |
Value (£): |
226,444
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| EPSRC Research Topic Classifications: |
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| EPSRC Industrial Sector Classifications: |
| Pharmaceuticals and Biotechnology |
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| Related Grants: |
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| Panel History: |
| Panel Date | Panel Name | Outcome |
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27 Jan 2010
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PDRF CDIP Interview Panel
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Announced
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17 Dec 2009
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PDRF CDIP Sift Panel
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Excluded
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Summary on Grant Application Form |
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In the biophysics and medicine field, researchers act as detectives working hard to unravel the mysteries surrounding cells. One type of spectroscopy in particular, Raman spectroscopy, has proven especially useful in providing detailed chemical composition of biological samples, constituting a sort of its chemical fingerprint. However the limited spatial resolution and sensitivity available is not sufficient to be able to resolve sub-cellular species. Furthermore, studies are normally performed on multiple cells grown or fixed on a surface which leaves a question about the influence of surface adhesion and cell to cell heterogeneities and interactions upon the biochemical state of the cells studied. Quantitative imaging and analysis of single normal and diseased cells is a challenging analytical goal. The principal objective of this project is to develop further a biophotonics technique that combine novel Raman spectroscopy methods with Optical Tweezers and apply this technique in obtaining time- and space-resolved information on biochemical and biophysical reactions inside single healthy and diseased living cells.In order to detect the distribution of assigned chemicals within a single selected cell, without physically touching it or needing to absorb it to a surface, it will be developed a new wide field spectral Raman imaging based on the use of Spatial Light Modulator (SLM) combined with an Optical Cell Rotator (OCR). This system will allow the acquisition of several spectra simultaneously from a large uniform area of interest of the optically trapped cell and incredibly will reduce the required acquisition time. So far, single living cells and subcellular compartments can be molecularly examined rapidly, non-invasively, without the use of external markers or fluorescent dyes and free from the interferences with the other cells.Although Raman spectroscopy is a useful technique to identify and quantify the biochemical composition of a cell, it can be severely limited in its applicability by sensitivity limits. Recently, numerous Raman based techniques have been developed in order to enhance the sensitivity of the basic technique. Among these, Surface-Enhanced Raman Scattering (SERS) is a process whereby the Raman scattering is enhanced, up to 14 orders of magnitude, when a Raman-active molecule is close to a metallic nanoparticle. Taking advantages from the major characteristics of both these new ultra-sensitive spectroscopic techniques (wide-field Raman imaging and SERS), it will be possible . to determine where assignment biochemical transformations occur on malignant cell membrane with spatial resolution of a few nanometers and sensitivity up to single molecules. All that has relevant implication for diagnostic and therapy of many diseases.Other important information can be obtained correlating such malignant biochemical transformations with the mechanical structure of normal and diseased cells. Recently, it has been demonstrated that mechanical stress can modify the protein expression in cancer cells membrane and consequently their susceptibility to the immune system detection. In particular, the project will focalize on the spectroscopic study of stress induced specific proteins in the various cellular compartments. This will require the development of micromanipulation apparatus (Optical Stretcher) that will be integrated with the ultra-sensitive spectroscopic techniques. Micromanipulation techniques will allow selecting a single cell and applying controlled mechanical stress (stretching) on it and ultra-sensitive spectroscopic techniques will allow the direct measurement of changes at molecular level (protein expression, cytoskeleton re-organization, etc.) that are directly related with mechanical stress.
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Key Findings |
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This information can now be found on Gateway to Research (GtR) http://gtr.rcuk.ac.uk
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Potential use in non-academic contexts |
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This information can now be found on Gateway to Research (GtR) http://gtr.rcuk.ac.uk
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Impacts |
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| Description |
This information can now be found on Gateway to Research (GtR) http://gtr.rcuk.ac.uk |
| Summary |
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| Date Materialised |
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Sectors submitted by the Researcher |
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This information can now be found on Gateway to Research (GtR) http://gtr.rcuk.ac.uk
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| Project URL: |
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| Further Information: |
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| Organisation Website: |
http://www.st-and.ac.uk |