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Details of Grant 

EPSRC Reference: EP/T033614/1
Title: Integrating confocal microscopy and cryo plasma FIB milling for tomography
Principal Investigator: Naismith, Professor JH
Other Investigators:
Dumoux, Dr M
Researcher Co-Investigators:
Project Partners:
Department: Research
Organisation: The Rosalind Franklin Institute
Scheme: Standard Research
Starts: 01 January 2020 Ends: 31 March 2021 Value (£): 1,200,000
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Summary on Grant Application Form
The Life Sciences sector forms a key part of the UK economy: it employs over 220,000 people, contributes significantly to

GDP and UK balance of trade, and is crucial for developing leading-edge treatments for patients. It is underpinned by the UK's world-leading research base in the health and life sciences. Many key research breakthroughs are, in turn, enabled by advances in engineering and physical sciences (EPS) research - which provide ever more sophisticated instrumentation and methods to support the study of living organisms (from microbes to plants, animals and the human body) and biological processes (including both disease pathology and drug action). R&D across all parts of this ecosystem - from fundamental understanding to applied research to product development - is crucial for the delivery of long-term economic growth and continued advances in agriculture, food security, healthcare and public health. Historic models of innovation have often been linear, involving a degree of serendipity. Disruptive technologies and scientific breakthroughs will be accelerated if physical scientists, engineers, life scientists and industry work together, and at scale. The Franklin has a focal point (Hub) at Harwell Science and Innovation Campus, linked to formal Spokes in leading HEIs across the UK, it will integrate complementary expertise from academia and industry to create a national centre of excellence for methods development at the convergence of the physical and life sciences.

It will create high-value jobs, protect and attract inward investment, and drive long-term growth; and contribute to the delivery of the Government's innovation, industrial and regional strategies.

The correlation of visible microscopy with electron imaging, is of course a critical goal of the RFI, it underpins imaging across scales. Thus for cells and tissue, the accurate targeting of the zone of interest to be milled is critical. Due to the crowded cellular environment, it is, as a practical matter, almost impossible to confirm the region of interest is within the EM volume during measurement, since the sample is damaged by electron dose. Correlative Light and EM (CLEM) methods are used to guide the milling and imaging. In broad terms fluorescent tag(s) are put onto the protein(s) of interest; this is extremely technically challenging and prone to error. We propose, in a collaboration with Thermo Fischer Scientific , to combine fluorescence imaging and plasma FIB milling in a single machine, thus transforming the workflow for tomography. This equipment has applications in both correlative imaging themes and in the mass spectrometry theme.

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