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 Liverpool, 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.
Polychromatic Optimization Protocol for Three Dimensional Immune and Structural Characterization of Whole-Mount Skin
Alfonso Schmidt Mery, Malaghan Institute of Medical Research, New Zealand
Skin is the first line of defence and the immune system’s biggest barrier for combating pathogens. Being able to accurately characterize and identify immune cell subtypes and structures in the skin under steady-state conditions, might provide a powerful tool for understanding the first immunological strategies and biological processes that occur in presence of pathogens.
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.
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.