Beating Hearts at High Resolution: Adaptive High Resolution Selective Plane Illumination Microscopy

Lead Research Organisation: Durham University
Department Name: Physics

Abstract

In this project we will film life as it develops in the heart by exploiting developments in optical microscopy, high speed imaging, and adaptive optics. The hearts will be in live, developing zebrafish and mouse embryos. This is extremely challenging as the living heart is large (on a microscopic scale), is embedded in the body of the animal (meaning it must be imaged at depth) and is beating, and therefore moving, at a rate of around 300 beats per minute. As well as providing a challenge to demonstrate the advanced methods we will develop the study of cardiac development is important both for investigating the development of the healthy heart and for determining the causes of heart problems.

Planned Impact

This multidisciplinary science project will have impact both in the core disciplines involved, physics, engineering and life science, but also into the commercial sector in areas from instrument development through to initial drug screening in the pharmaceutical industry. We will publish high quality papers in leading journals in the areas of optics, active computer control, novel microscopy and cell and developmental biology All of the applicants have an excellent track record in interdisciplinary research and publication and are frequently invited to speak at international conferences. We believe that this proposal will thus have academic impact in many scientific fields, which will enhance its scientific impact and the project will also aim to stimulate multidisciplinary research. Physical Science Context The instruments developed in this project will for the first time incorporate adaptive optics for sample induced aberration correction in a SPIM system, and in more general terms the use of a true closed loop AO system incorporating a real time wavefront sensor. The knowledge gained from this work will have applications in advancing the incorporation of AO in microscopy in other, more traditional, beam scanned imaging systems leading to significantly improved in depth imaging. Biological Context Cardiac malformation is the leading cause of illness and death in children in the western world, with almost 1% of babies born with a significant congenital heart defect. Moreover, it is becoming clear that predisposition to some forms of adult heart disease may result from subtle abnormalities in heart development, that were asymptomatic at birth, but which progressively impact negatively on cardiac function. If we are to understand the causes, and ultimately prevent these malformations, it is essential to understand the fundamental processes that form and remodel the developing vertebrate heart. Whereas there has been a great deal of progress in recent years in understanding the genetic networks that regulate heart development, the cellular processes that these control, and the non-genetic factors that modulate these cellular behaviours (such as haemodynamic forces as blood flows through the forming heart), remain unclear. Furthermore, bioinformatic approaches and genome wide sequencing approaches continue to provide candidate mutations within genes that may have clinical relevance. The ability to assess the significance of these mutations on cardiac form and function in tractable animal models would greatly improve our understanding of disease mechanisms. Making the Technology Available In a commercial context the Durham group are in conversation with several microscopy companies about the incorporation of AO into commercially available imaging systems and the team, through Prof Girkin, have already helped to install AO systems on several beam scanned systems around the world that are now in routine use within biological laboratories. Thus there is a tried and proven route in helping such instrument developments to be applied in the end user's laboratories. As the project develops consideration will be given to the potential for the protection of the intellectual property emerging from the project and an agreement will be put in place between the two Universities to cover this aspect of the work. Broader Impact During the project we will also arrange at least one multidisciplinary meeting dedicated to the subject of imaging and development biology. Towards the end of the project we will arrange a similar meeting with the aim of stimulating further discussion and research in the area, with a strong emphasis on demonstrating how the integration of experimental optics, computer image analysis and advance biology can lead to significant breakthroughs for the benefit of all.

Publications

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Bourgenot C (2013) Comparison of closed loop and sensorless adaptive optics in widefield optical microscopy in Journal of the European Optical Society: Rapid Publications

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Bourgenot C (2012) 3D adaptive optics in a light sheet microscope. in Optics express

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Bourgenot, C. And Saunter, C. D. And Girkin, J. M. And Love, G. D. (2011) Adaptive Optics for Wide-field Microscopy

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Bourgenot, C. And Saunter, C. D. And Love, G. D. And Girkin, J. M. (2013) Comparison of closed loop and sensorless adaptive optics in widefield optical microscopy in {JOURNAL OF THE EUROPEAN OPTICAL SOCIETY-RAPID PUBLICATIONS}

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Bourgenot, Cyril And Saunter, Christopher D. And Taylor, Jonathan M. And Girkin, John M. And Love, Gordon D. (2012) 3D adaptive optics in a light sheet microscope in {OPTICS EXPRESS}

 
Description We have developed a new type of microscope which allows us to film the development of organs in an animal as the develop.
Exploitation Route We have build a second microscope to be used by medical researchers at Edinburgh University.
Sectors Healthcare,Pharmaceuticals and Medical Biotechnology

URL https://www.dur.ac.uk/cfai/biophotonics/advancedmicroscopy/spim/
 
Description We developed a selective plane illumination microscope - incorporating adaptive optics for high resolution imaging. We also developed a new type of image synchronisation system which allowed us to record volumetric images of the beating heart of embryonic zebrafish.
First Year Of Impact 2004
Sector Healthcare,Pharmaceuticals and Medical Biotechnology
Impact Types Societal

 
Description SPIM Collaboration with Edinburgh 
Organisation University of Edinburgh
Department School of Geosciences Edinburgh
Country United Kingdom 
Sector Academic/University 
PI Contribution We have built a specialised microscope for the team at Edinburgh to further their research.
Collaborator Contribution We have built a specialised microscope for the team at Edinburgh to further their research.
Impact Papers
Start Year 2012
 
Description SPIM with Edinburgh 
Organisation University of Edinburgh
Department Centre for Integrative Physiology
Country United Kingdom 
Sector Academic/University 
PI Contribution We constructed a SPIM microscope for the team at Edinburgh
Collaborator Contribution They continue to use the microscope for research
Impact See papers
Start Year 2012
 
Description British Association for Science 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact We built a large model microscope system which we display at the British Association Festival of Science in Newcastle in 2013
Year(s) Of Engagement Activity 2013
URL http://www.newcastlegateshead.com/whats-on/british-science-festival-p687051
 
Description Jeremiah Horrocks Public Lecture 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact John Girkin gave the Jeremiah Horrocks public lecture at the University of Central Lancashire.
Year(s) Of Engagement Activity 2013
URL https://www.youtube.com/watch?v=LmKYST6O2M8