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
Bourgenot C
(2013)
Light sheet adaptive optics microscope for 3D live imaging
Bourgenot C
(2012)
3D adaptive optics in a light sheet microscope.
in Optics express
Bourgenot C
(2013)
Comparison of closed loop and sensorless adaptive optics in widefield optical microscopy
in Journal of the European Optical Society: Rapid Publications
Bourgenot C
(2011)
Adaptive optics for wide-field microscopy
Bourgenot, C. And Saunter, C. D. And Girkin, J. M. And Love, G. D.
(2011)
Adaptive Optics for Wide-field Microscopy
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}
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}
Buckley C
(2017)
Precise spatio-temporal control of rapid optogenetic cell ablation with mem-KillerRed in Zebrafish.
in Scientific reports
Chalmers S
(2010)
In Smooth Muscle, Mitochondrial Movement is Restricted in Native Cells and Unrestricted and Trafficked When Cells are in Culture
in Biophysical Journal
Love G
(2014)
Handbook of Liquid Crystals
Matrone, Gianfranco And Taylor, Jonathan M. And Wilson, Kathryn S. And Baily, James And Love, Gordon D. And Girkin, John M. And Mullins, John J. And Tucker, Carl S. And Denvir, Martin A.
(2013)
Laser-targeted ablation of the zebrafish embryonic ventricle: A novel model of cardiac injury and repair
in {INTERNATIONAL JOURNAL OF CARDIOLOGY}
Mitchell TJ
(2014)
Quantitative high dynamic range beam profiling for fluorescence microscopy.
in The Review of scientific instruments
Silburn, Scott A. And Saunter, Christopher D. And Girkin, John M. And Love, Gordon D.
(2011)
Multidepth, multiparticle tracking for active microrheology using a smart camera
in {REVIEW OF SCIENTIFIC INSTRUMENTS}
Taylor J
(2012)
High-resolution 3D optical microscopy inside the beating zebrafish heart using prospective optical gating
in Biomedical Optics Express
Taylor JM
(2011)
Real-time optical gating for three-dimensional beating heart imaging.
in Journal of biomedical optics
Taylor JM
(2014)
Optically gated beating-heart imaging.
in Frontiers in physiology
Taylor, J. M. And Saunter, C. D. And Chaudhry, B. And Henderson, D. J. And Love, G. D. And Girkin, J. M.
(2011)
Heart Synchronization for SPIM Microscopy of Living Zebra Fish
Taylor, J. M. And Saunter, C. D. And Love, G. D. And Girkin, J. M.
(2013)
High-resolution time-resolved 3D optical microscopy inside the beating zebrafish heart using prospective optical gating
Taylor, J. M. And Wong, L. Y. And Bain, C. D. And Love, G. D.
(2008)
Emergent properties in optically bound matter
in {OPTICS EXPRESS}
Taylor, Jonathan M. And Girkin, John M. And Love, Gordon D.
(2012)
High-resolution 3D optical microscopy inside the beating zebrafish heart using prospective optical gating
in {BIOMEDICAL OPTICS EXPRESS}
Taylor, Jonathan M. And Saunter, Christopher D. And Love, Gordon D. And Girkin, John M. And Henderson, Deborah J. And Chaudhry, Bill
(2011)
Real-time optical gating for three-dimensional beating heart imaging
in {JOURNAL OF BIOMEDICAL OPTICS}
Thalhammer G
(2013)
Speeding up liquid crystal SLMs using overdrive with phase change reduction.
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 |