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

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.

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.