Proposal for a Talos L120C G2 Transmission Electron Microscope

Lead Research Organisation: The Francis Crick Institute
Department Name: Research

Abstract

The Crick is a partnership between the Medical Research Council, Cancer Research UK, the Wellcome Trust and three leading universities: UCL (University College London), Imperial College London and King's College London. The Crick aspires to be one of the world's leading medical research institutes. The Crick achieves operational and research efficiencies and economies-of-scale through centralised facilities and functions, known as Science Operations, that provide all researchers at the Crick, irrespective of affiliation, with access to cutting-edge equipment, animals for research and laboratory enabling functions such as the Electron Microscopy Science Technology Platform (EM STP).
Electron microscopy is a fundamental tool in bioscience research. It is used to image the structure of proteins, pathogens, cells and tissues, to help us understand the biological processes of normal development, disease states, and the effect of therapies.
This proposal is for a new transmission electron microscope that will be used for state-of-the-art imaging experiments for 60-80 research projects ongoing at the Crick. The microscope has a range of additional features, including automated image stitching to allow us to view large samples and digitally zoom into features of interest (like 'google maps'); faster, more automated, three-dimensional imaging of samples using a technique called electron tomography; and a cryo-stage for imaging samples encased in ice, as close to their living state as possible.

Technical Summary

The Crick aspires to be one of the world's leading medical research institutes. It achieves research efficiencies through centralised facilities and functions such as the Electron Microscopy Scientific Technology Platform. A BBSRC-funded transmission electron microscope would be placed in the EM STP and made available to 100+ research groups at the Crick, and their collaborators. This application is being made by twelve Crick Group Leaders with Lucy Collinson, Head of the EM STP. These diverse projects (illustrative of the breadth of research at the Crick), are united by the need for high resolution imaging.
- Pachnis: Investigating whether her4.1+ glial cells are an elusive population of enteric neural stem cells.
- Boulton: Mechanics of joint molecule formation during homologous recombination.
- Gutierrez: Cellular mechanisms of Mycobacterium tuberculosis replication in autophagosomes of endothelial cells and in necrotic macrophages.
- Lovell-Badge: Identification and characterization of NG2-glia and their interactions in the median eminence of mouse hypothalami.
- Ultanir: Ultrastructural features of ependymoma-like Lats1/2 knockout tumours.
- Tooze: Resolution of membrane contact sites required for autophagosome formation.
- Parker: Unravelling the role of stress-granule-like structures in protection against mitotic failure.
- Sahai: Ultrastructure of pathological cell-cell contacts between epithelial cells and fibroblasts and the complex membrane interface that enables transcytosis between cells.
- Gould: Mapping the accumulation of lipid droplets in a Drosophila renal models of metabolic syndromes.
- Deu: Chemical and genetic interrogation of novel proteolytic pathways in Apicoplexan parasites.
- Nurse: Role of membrane flow between the nuclear envelope and endoplasmic reticulum in nuclear size homeostasis in fission yeast.
- Blackman: Morphology, biogenesis and molecular makeup of the malaria parasite Plasmodium falciparum secretory organelles.

Planned Impact

The Crick aspires to be one of the world's leading medical research institutes. Its multidisciplinary approach, an emphasis on practical application of research and its links with academia, industry and the public sector will speed up the translation of discoveries made in the laboratory into treatments for disease. We fully expect outcomes from the Talos L120C G2 TEM and its use by researchers across the Crick to have a pathway to impact through academic, economic and societal impacts.
Academic impact:
- Publication via various media including papers and conference presentations;
- Knowledge transfer and education, especially with respect to PhD students and early-career researchers / post-doctoral research associates.
Specifically, the EM STP has a track-record in broadening impact of equipment awards. As part of the successful BBSRC/MRC/EPSRC Next Generation Optical Microscopy Award in 2013, the EM STP committed to make a proportion of time on their Serial Block Face SEM and Integrated Light and SEM openly accessible (and free at point of access) to groups outside the core-funded CRUK London Research Institute (one of the founder institutes of the Crick and the previous home of the EM STP). This award resulted in seven technology development publications [1-6], and 10 primary bioscience papers to-date from the open access commitment [7-16], with another four manuscripts submitted or in revision.
The impact of this award was broader than even we envisaged, thanks to the open access commitment but also to the training that was given to each of our external collaborators in sample preparation and correlative microscopy, to enable them to prepare samples remotely for imaging in the EM STP. Indeed, our collaborations from this award stretched from Aberdeen, Cambridge and Bristol to Stellenbosch, Leiden and Massachusetts. This proof-of-concept worked so well that we propose to formalise our collaboration with the BBSRC-funded Babraham Institute by making a proportion of the time on the new microscope open access for their research groups (at least 10%, depending on their needs). We have supported the Babraham over the last four years in their testing and installation of a Focused Ion Beam SEM, in data collection for projects with the Ktistakis and Florey labs as part of our open access commitment, and in training for correlative microscopy. The new TEM would complement three-dimensional data from the Babraham's new FIB SEM with high resolution membrane bilayer data where the research requires it (see letter of support from Professor Michael Wakelam, Director of the Babraham Institute).
Economic and societal impact:
- The Crick, its founding partners (the Medical Research Council, Cancer Research UK, the Wellcome Trust, UCL (University College London), Imperial College London and King's College London), and legacy research institutes at Mill Hill (MRC), Lincoln's Inn Fields and Clare Hall (CRUK) all have strong track records for converting research and discoveries into economic and societal benefit;
- The Crick organisation includes well-funded and specific groups responsible for Translation, Clinical Research, Public Engagement and Education, thus we are confident that research outcomes from the BBSRC funded Talos L120C G2 TEM will convert into economic and societal benefits. It is worth emphasising that the microscope will be housed in the Crick's Electron Microscopy STP, and hence be available to 100+ Crick research groups as well as our collaborators in partner institutions, such that the pathways to impact are many and varied.

Publications

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