We have launched a brief survey examining the needs in the Life Sciences around the use or potential utility of optical (fluorescence) microscopies. This takes ~ 6 minutes to complete and gives you an opportunity to stay in touch or engaged directly with the new developments of the newly launched UKRI Future Leader Fellowship within the Applied Biophotonics Group. Whether you are an experienced microscopist or never had the opportunity to use optical imaging, can such microscopies add value to your research or day-to-day work? If so, what holds you back? If you work within any area of Life Sciences, we would like to hear about your experiences. Please take the survey and let us know.
1st of May – We have officially moved our research operations to the Department of Molecular Biology & Biotechnology in our new home institute, the University of Sheffield. We will be based in the famous Firth Court, working closely with a wide range of interdisciplinary research groups and the IMAGINE imaging consortium.
The UK is currently in the midst of the lockdown due to the ongoing COVID-19 pandemic. Normal operations will therefore begin in the coming months. There will be a number of professional and study opportunities within our team coming very soon. So watch this space!
The principal investigator of the Nanoscale Microscopy Group, Dr Izzy Jayasinghe, has been awarded a UKRI Future Leader Fellowship to build a new portable imaging technology which will allow scientists, medical doctors, conservationists and industrial parties to visualise the smallest building blocks of any biological sample from any location.
The UK and other governments are currently making urgent investments into understanding the role of molecules and cells in some of the biggest challenges of today’s society which include the effects of climate change on food sources and lifestyle effects on major human diseases and ageing. Despite this urgency, we remain unable to visualise the relevant genes, proteins and cellular components in their natural environments or the geographical locations of the problem. Frustratingly, the technology for visualising such minute structures exists. It is called ‘super-resolution microscopy’ and we even hailed its invention with a Nobel Prize in Chemistry in 2014. However, it has remained beyond the reach of field scientists and clinicians because it has always relied upon specialist skill for its operations and expensive and bulky equipment for its implementation.
In this fellowship, we will use a radically new approach to make super-resolution microscopy portable, cheap and easy to use. We will harness a novel chemical reaction called ‘Expansion Microscopy’ which we have refined and mastered over the last three years (read more about our recent paper about it here). This method allows one to physically inflate a desired feature of a sample, for example a patient biopsy or a small organism, by over a 1000-fold in volume. We will build a set of chemical, biological and physical tools which allows this method to reveal minute cellular details in tissues or organism which were previously too small to be visualised with traditional, laboratory-based, optical microscopes. These developments will be carried out with a view to assemble a miniature super-resolution microscope that is both affordable and portable beyond the laser lab.
To refine and ensure that this device delivers this claimed imaging capability, we will carry out case studies in partnership with experts whose samples are collected outside of the academic laboratory (in Phase II). They include a field scientist who will use it to examine young sea urchins in the UK coast, doctors and sports scientists who will screen for the fine structure of needle biopsies taken in the clinic from human patients, and a member of the Worms in Space programme who will use it to remotely study the effect ‘zero gravity’ on the ageing of microscopic worms sent between earth and the international space station.
Our expectation is that by making super-resolution available beyond the laboratory, one unlocks the benefits of rapid visualisation of sub-cellular structures which underpin the life processes and pathology at a new spacial scale. For field scientists, it would accelerate research programmes; sample collection and high-end microscopic analyses would no longer be mutually exclusive processes. In the clinic, this could unlock faster decision-making.
The fellowship allows us to work more closely with two important industrial partners who have supported us over the last few years, Badrilla Ltd and Cairn Research.
We demonstrate the ability to exploit in-plane resolution of ~ 15 nm and axial resolution of ~ 35 nm by combining X10 Expansion Microscopy with Airyscan 3D imaging.
Expansion Microscopy is the newest of super-resolution imaging methods which allows finer details of samples to be visualised with relatively conventional fluorescence imaging techniques by physically expanding the sample. This is achieved by embedding the samples in an acrylamide hydrogel matrix, crosslinking the fluorescent probes, enzymatically clearing the sample and then osmotically swelling the hydrogel (this paper demonstrates ~ 1000-fold volume expansion).
The enhanced three-dimensional resolution achieved by combining this with Airyscan microscopy (hence, the name Enhanced Expansion Microscopy, or EExM) is better suited than conventional implementations of localisation microscopies, particularly for imaging cell interiors and ultrastructures in cell types with 3D complexity, more so than some of the more popular super-resolution techniques. We demonstrate this by imaging cytoskeletal alpha-actinin lattices and RyR nanodomains in ventricular cardiomyocytes. We go a step further to show how this single-channel resolution can reveal dispersed RyR array structures and the altered single-channel phosphorylation patterns which coincide with the fatal heart pathology – right ventricular failure. To better-understand the functional implications, we have teamed up with Dr Michael Colman (http://physicsoftheheart.com/) to simulate the local calcium signalling events based on the experimentally-mapped RyRs.
This is the first research paper for Tom Sheard, and marks the first home-publication for the Nanoscale Microscopy Group together with a multi-lateral collaboration. Well done, everyone! This work was funded by the MRC DiMeN and Wellcome Trust Seed Award.
On the 12 October, Miriam ran an event funded by The Physiological Society to promote Physiology Friday at The University of Leeds. The aim of this outreach event was to raise awareness of how technological advancements in microscopy are improving our understanding of how human physiology can change within health and disease at the cellular level, specifically related to the heart.
Within research, the knowledge of our own physiology in health and disease has developed in part from the advancement in microscopy, which has enabled the visualisation of single proteins. This advancement can be likened to the difference between an individual who is legally blind compared to someone who has 20/20 vision. Our event utilised visual impairment glasses which distorted an individual’s vision. Participants were asked how well they could resolve the outline of a structure in the absence and presence of the glasses to convey the significance of the development which has occurred within microscopy. Supplementary posters led individuals through the physiological relevance of this technological advancement, with images detailing the single protein structure within the heart. With the use of an anatomical heart model, the function and structure of the heart within health and disease was explained. We communicated how current research is using a super-resolution version of microscopy to characterise how a change in function during heart failure can be due in part to a change in the protein structure within a heart cell. To help convey the scale at which the cellular remodelling within heart failure is occurring, real-life examples were used. For example, if a person was tall enough to reach a plane flying at 30,000 feet then the proteins that we are imaging would still be smaller than the width of that person’s hair. Smartphone microscopes were available to allow individuals to experience first-hand the ability that a microscope has to study an object at a more detailed scale compared to what can be seen by the human eye alone.
Over the course of the day a dialogue was created with students from all disciplines. This sparked a variety of questions on the topic of physiology; what is a heart attack? What happens to the proteins within your heart during heart failure? The event even led to one student exclaiming that “microscopes are so cool”, with another stating “I learned how much I need to take care of my body”.
Many thanks to The Physiological Society who funded and supported this event – now to start planning the next one!
In June 2018 Miriam attended a Public Engagement Training Day which was run by The Physiological Society. Building upon the skills learnt at this event Miriam gained funding from the Society to organise and run an event at The University of Leeds to promote Physiology Friday. Her reflections upon this training day can be read on Page 47 in the Winter 2018 Issue of Physiology News which can be accessed here.
In October 2018 I flew across the world to the laboratory of Dr David Crossman at the University of Auckland in New Zealand. The goal: to study pathological remodelling in human heart biopsies from patients with idiopathic dilated cardiomyopathy (IDCM), using the super-resolution imaging technique expansion microscopy (ExM).
Our interest lies in seeing whether nanodomain remodelling observed in a rat model of heart failure in Leeds, including reorganisation of the internal calcium compartments and functional modification to calcium-handling proteins, is also present in end-stage human heart failure. Understanding the mechanisms of remodelling is one of the first steps towards investigating whether they can be targeted for preventative therapies.
ExM is novel imaging technique, enabling super-resolution imaging by spatially separating fluorophores within a swellable hydrogel. The compatibility of ExM gels with standard microscopes enables greater imaging depth and improved axial resolution over competing super-resolution techniques. ExM therefore provides a practical tool to observe remodelling within dyadic calcium release clusters. I was responsible for starting ExM experiments from scratch in a new lab across the world, requiring efficient independent work to obtain meaningful data in the space of just 4 weeks.
It was fantastic to take this journey and work in a laboratory that is home to a strong consortium of leading cardiovascular researchers. In my final week I gave a 30-minute seminar, in which I presented work to the physiology department and the wider bio-imaging facility. This allowed me to reach an international audience and receive valuable feedback on the progression of my research.
Many thanks to the MRC and DiMeN flexible fund grant which made this trip possible, and special thanks to David Crossman for welcoming me into his lab.