Details of Grant 

EPSRC Reference:	EP/H03238X/1
Title:	Development of Oblique Plane Microscopy for Biomedical Applications
Principal Investigator:	Dunsby, Dr CW
Other Investigators:
Researcher Co-Investigators:
Project Partners:
Department:	Physics
Organisation:	Imperial College London
Scheme:	First Grant - Revised 2009
Starts:	01 June 2010	Ends:	31 July 2011	Value (£):	101,196
EPSRC Research Topic Classifications:	Med.Instrument.Device& Equip.
EPSRC Industrial Sector Classifications:	Healthcare
Related Grants:
Panel History:	Panel Date Panel Name Outcome 09 Feb 2010 Materials, Mechanical & Medical Engineering Panel Announced
Summary on Grant Application Form
Conventional optical microscopy uses visible light to provide high resolution (>~200 nm) images for a huge range of applications from industrial inspection of electronic devices to biomedical imaging of cells and tissue. Fluorescence microscopy is an extension of optical microscopy that has become a standard tool for biologists who label their specimens with fluorescent molecules to report the locations of specific proteins, or to study cellular processes, e.g. by imaging proteins interacting.Many microscopy applications demand optically sectioned imaging, which provides an image of only a single thin (~1 micron) slice through the sample. Optical sectioning improves image contrast by reducing the 'blur' from out-of-focus planes that is evident in conventional microscopy and provides the ability to produce 3D images from stacks of 2D optically sectioned images. Normally, optically sectioned imaging is implemented using expensive laser scanning confocal microscope systems (typically >150k), which can be considered as a gold standard . While these microscopes provide high resolution 3D images, they typically require 10's of seconds to acquire a 3D fluorescence intensity image. Also, when imaging at higher speeds, the illumination used in confocal microscopes can cause light induced damage (photodamage) to biological samples.Several alternative optical sectioning microscopy techniques have been developed that generally address some of the disadvantages of confocal microscopy, but which inevitably present a compromise elsewhere. One recently developed alternative is selective plane illumination microscopy (SPIM), which provides rapid optically sectioned imaging and with very low exposure of the sample to illumination light, and which is particularly advantageous when imaging live biological specimens. However, a major disadvantage of SPIM is that it cannot be implemented on the standard fluorescence microscopes that are used widely in biomedical research.This project will develop a new optically sectioning microscope technology invented by the applicant called Oblique Plane Microscopy (OPM, patent filed July 2008). OPM is conceptually similar to SPIM but it can be implemented on standard fluorescence microscopes and applied to image samples prepared on standard microscope slides or the standard cell culture dishes or multiwell plates that are used by the vast majority of biologists. As for SPIM, the image acquisition rate of OPM is only limited by the speed of the CCD camera used (e.g. 1000 frames per second) and OPM subjects the specimen to a minimal light exposure. This project will apply OPM to two different biological applications. The first will image isolated beating heart muscle cells to measure and quantify the propagation of calcium waves and to image small rapid changes in calcium concentration known as 'calcium sparks'. OPM will provide high speed optically sectioned fluorescence detection to image spark events and will also be used for time-lapse 3D imaging of calcium wave propagation within individual heart muscle cells. The second biological application will be to image small (~50 micron) fluorescently labelled zebra fish embryos. The results obtained on these biological samples will demonstrate the utility of OPM and provide preliminary data for future interdisciplinary research projects.A key advantage of OPM is the potential for high-speed high-throughput automated 3D imaging, e.g. in multi-well plates used in biological and drug-discovery assays. This project aims to demonstrate this potential, which is important for commercial exploitation. A further route to commercialisation of OPM could be as a 'bolt-on unit' to a conventional fluorescence microscope that could provide optically sectioned imaging at significantly lower cost than a laser scanning confocal microscope.
Key Findings
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