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Details of Grant 

EPSRC Reference: EP/F042647/1
Title: Fast remote focusing for three-dimensional imaging of biological tissues.
Principal Investigator: Botcherby, Dr EJ
Other Investigators:
Researcher Co-Investigators:
Project Partners:
Ecole Polytechnique
Department: Engineering Science
Organisation: University of Oxford
Scheme: Postdoc Research Fellowship
Starts: 01 September 2008 Ends: 02 December 2010 Value (£): 245,293
EPSRC Research Topic Classifications:
Cells Med.Instrument.Device& Equip.
EPSRC Industrial Sector Classifications:
Healthcare
Related Grants:
Panel History:
Panel DatePanel NameOutcome
10 Mar 2008 LSI Postdoctoral Fellowships Interview Panel 2008 Announced
11 Feb 2008 LSI Postdoctoral Research Fellowships 2008 InvitedForInterview
Summary on Grant Application Form
The multiphoton microscope is a powerful imaging tool that is widely used across the biological sciences. The strength of this system, compared with the conventional optical microscope, lies in its ability to image specimen structures at high resolution in three-dimensions rather than two-dimensions. Typically, this is achieved by refocusing the microscope and acquiring a series of two dimensional images at a number of different specimen depths. The resulting image stack can then be manipulated by computer algorithms in a number of different ways to reveal a wealth of information about the object structure. Recently, it has become of interest to collect these three dimensional image stacks at high speed in order to observe the dynamic behaviour of biological specimens. As a result, technological advances have been made to improve image acquisition speeds.The problem is that although a single in-focus image can be obtained very quickly the real bottleneck in three-dimensional data acquisition is the process of refocusing the microscope to successive image planes. For fundamental optical reasons, refocusing must be carried out by physically changing the distance between the objective lens and specimen, which is problematic for two reasons. First, this process is generally slow and second it can lead to undesirable specimen agitation.In order to alleviate these restrictions, I have recently developed a new system architecture that permits refocusing to be carried out at far superior speeds than current technology will allow. Furthermore, refocusing is carried out remotely from the specimen, which simulates a more natural environment in which to perform investigations. During this research project, I propose to implement this new technique on a two photon microscope to permit a number of unique imaging modalities. For instance, this system will be able to acquire images directly from a curved surface at high speed. Previously, such information could only be derived from a three dimensional image stack, which, due to the aforementioned limitations, would have lower temporal resolution. This is particularly of interest in applications where the dynamic behaviour of cell movements is limited to take place over a well defined surface that is not flat. Furthermore, I propose to apply this technique of refocusing to a microscope system that already exists at the Laboratory of Optics and Biosciences in Paris. This specialized system acquires three-dimensional data with a number of different imaging modes and has already been used to investigate the early stages of embryo development, where cell movements are mainly localized around the embryo surface. As a result of this new refocusing strategy, such biological problems will be investigated further and better insight gained.
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