Below you can find out the work that is being undertaken by our current RMS Diploma students:
Developing protocols and training materials for the live imaging of Drosophila via lightsheet and multiphoton microscopy
Jennifer Adcott, University of Manchester, UK
Lightsheet and multiphoton microscopy have been widely used to reduce photo-damage. Here these systems will be used and compared for the imaging of live Drosophila using established cancer model fly lines. A protocol will be established for sample preparation, image acquisition, through to image processing and analysis. Suitability for each technique will be compared for addressing specific questions where live imaging of small model organisms or ex vivo tissues is required. The aim will be to publish these results as a methodology paper with JoVE video protocol.
The Characterisation of Speciality Fibres Utilising 3-D SEM Constructed Images
Michael Brookes, University of Leeds, UK
Distinguishing expensive speciality animal fibres such as cashmere and alpaca from cheaper fibres such as wool is an important activity. Current techniques, such as ISO 17751-2, measure fibre diameter utilising an SEM in 2 dimensions at high vacuum and susceptible to viewing errors. The study will use 3-D imaging techniques to measure and characterise speciality fibres and compare the results with that of current techniques.
Testing, optimising and implementing cryo-sample preparation and imaging techniques to optimise sample quality and acquisition efficiency in single particle analysis
Adam Costin, University of Sydney, Australia
With the rise of cryo-EM as a powerful tool for solving protein structures there has been an influx of new researchers learning the technique. Sample optimisation for cryo-EM remains slow and highly inconsistent. This study will aim to create a workflow that will systematically optimise sample preparation much faster for new users. This will be done through a comprehensive assessment of current techniques and then testing these within the context of an EM Facility. A way of simply incorporating tomography into the process will also be developed, as this is a powerful way of assessing ice thickness and protein structure.
Electron Microscopy Study of Precipitates in Energy Generation Steels
Samuel Davis, Loughborough University, UK
Using a multimodal approach to compare precipitate imaging techniques of power generation steels. Both Scanning Electron and Focused Ion Beam (FIB) systems will be used to collect images, employing a combination of modern detectors, and optimised operating parameters for each system. These will be compared to benchmark examples cited in the literature with a view to improving upon the currently referenced standards.
Development of Quantitative Single Molecule RNA Imaging in Plant Crop Species
Susan Duncan, John Innes Centre, UK
Quantitative single molecule RNA imaging is highly complementary to next generation RNA sequencing approaches. Although lower throughput, imaging has an advantage over sequencing as it reveals subcellular RNA localisation patterns that can have significant consequences for gene regulation. Despite advances in RNA imaging in many other model organisms, for plant biology this approach is currently limited to a few tissues in the model plant Arabidopsis thaliana. The project proposed for this Diploma aims to expand this imaging approach to a wider range of tissues in commercially important crop plants including wheat, sugar beet and oil seed rape.
Investigation of the dynamic localisation and dimerisation of ATM kinase
Rebecca Gascoyne, University of Birmingham, UK
Ataxia-Telangiectasia Mutated (ATM) is the central regulator of the DNA damage response in cells. ATM is mainly located in the nucleus or cytosol but has also been found at synapses – and the significance of this is unknown. It is known that ATM exists both in a monomeric and dimeric form but is unknown what significance the localisation and dimerisation has. Using confocal and super-resolution microscopy, the localisation and dimerisation status of ATM will be observed before and after DNA damage in neuronal and non-neuronal cells, to observe its pattern of behaviour and speculate as to its significance in the DNA damage response.
The development of teaching materials for the use of scanning electron microscopy for secondary school education
Ramona Szalczinger, Tonbridge School, UK
My study aims to help lift practical scanning electron microscopy into secondary school education, and thus encourage STEM learning by providing teaching staff with teaching materials and lesson notes to match the national curriculum.
Application of Deep Learning Tools for Imaging Cytometry to Enable Label Free Characterisation of Manufactured Red Blood Cells
Pamela Twist, Safi Biotherapeutics, UK
In the manufacture of red blood cells (RBCs), it is crucial to track the maturation of cells in the bioreactor from a hematopoietic progenitor cell to a mature RBC. This can be achieved by fluorescent staining of maturation markers. Using imaging cytometry, we can collect the fluorescent signal of cells along with their corresponding brightfield image that captures morphological changes of maturing RBCs. My study aims to investigate if we can apply deep learning tools to our imaging cytometry data, so we can characterise the stage of maturation of manufactured RBCs in a label free manner.
Rapid and Non-Destructive Analysis of fresh or Formalin-Fixed Tumor Biopsies Using Fluorescence Microscopy and Artificial Intelligence for Image Reconstruction
Krunoslav Vinicki, Digicyte Ltd, Croatia
The study aims to develop and validate a new approach in microscopy for the rapid and
non-destructive analysis of fresh tumor biopsy samples. With this method I'm planning to address a longstanding need in pathology for real-time, on-site biopsy examination while preserving the tissue for further analysis. This could expedite the initiation of targeted therapy and reduce the need for repeat sampling due to insufficient material.
Development of image analysis routines to study the dynamics of centrosomes and centrioles in Drosophila
Alan Wainman, University of Oxford, UK
Imaging has evolved significantly in the last few years, and no longer involves just the capture of beautiful images, but also the generation of large datasets that must be quantified and analysed. The main goal of this proposal is to develop my image analysis skills, initially through the attendance of courses and subsequently through the development of scripts for ongoing research. These skills will ultimately benefit not only my own research but also the support I provide to users of my imaging facility. I also hope to record my learning experience to help others hoping to develop their own image analysis skills.
Assessing the distribution and localisation of the Human Silencing Hub (HuSH) in embryonic stem cell differentiation
Timothy Young, University of Cambridge, UK
Our genome is under constant threat from invasion by mobile genetic elements, including viruses (such as HIV) and retrotransposons (Line 1) which make up 17% of our genome. The Lehner group previously described the Human Silencing Hub (HuSH) complex and showed how it silences genome invaders through chromatin (H3K9me3) modification. HuSH is comprised of three core component proteins: TASOR, MPP8 and Periphilin. The intracellular localisation of these proteins is poorly characterised, particularly their intranuclear location and role in stem cell differentiation. I intend to utilise Super Resolution and Single Molecule Localisation Microscopy (SMLM) techniques to elucidate the HuSH intranuclear location in human cell lines and at different timepoints during mouse and human stem cell differentiation.