Preprint outlining the utility of peptide ligands against RyR2 as potential super-resolution imaging probes

Our latest pre-print is now online, where we evaluated two peptide ligands of the cardiac ryanodine receptor (RyR) as super-resolution imaging probes. The research was conducted in collaboration with one of our industrial partners, Badrilla Ltd., during the industrial internship of iCASE PhD student Tom Sheard funded through the DiMeN DTP.

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The peptides used were the scorpion venom toxin imperacalcin, and CPVT-mimicking DPc10. We evaluated the specificity of the fluorescent peptide conjugates for the target RyR2 in fluorescent imaging experiments, in terms of colocalisation with RyR2 immunolabelling, with confocal and expansion microscopy.

We also performed structural work looking at the DPc10 binding site on RyR2 atomic models to learn more about the peptide’s mechanism of disturbance (DPc10 binding to RyR2 causes domain unzipping and destabilises the channel).

Thanks to the host company Badrilla, the DiMeN DTP for funding Tom’s industrial placement, and the other authors for help with the research.

Access the pre-print in bioRxiv at: https://www.biorxiv.org/content/10.1101/2020.06.26.131854v1.

Feedback on all contents of the work is welcomed.

Follow Tom on twitter : @tmdsheard. https://twitter.com/TMDSheard

New book chapter detailing EExM in Methods in Cell Biology

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How we prepared EExM samples

Our new book chapter has been published, detailing the enhanced expansion microscopy technique (EExM). The protocol offers a route to visualising cellular ultrastructures at a resolution of 15 nm in-plane (and ~ 35 nm axially) with a hydrogel-based molecular-scale imprint of the fluorescence pattern.

We detail how we used EExM to explore structural and biochemical remodelling in cardiomyocytes. The chapter includes a step-by-step protocol, reagent list and troubleshooting tips.

Access the paper published in Methods in Cell Biology at: https://sciencedirect.com/science/article/pii/S0091679X20301084.

Any difficulties accessing the article please contact the Tom Sheard via email at: bs11ts[at]leeds.ac.uk

This is the first book chapter for PhD student Tom Sheard (@tmdsheard on Twitter).

 

Optical microscopy bottlenecks survey

bottleneck-910050_1280We 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.

 

We have arrived in Sheffield

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!

UK Research & Innovation Future Leader Fellowship awarded to develop a compact super-resolution microscopy technology

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.

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A fluorescence micrograph of a sea-urchin embryo which will be the subject of a field-trial for the portable super-resolution microscopy technology, currently under development. Image credit: Dr Andrea Gaion

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.