Current Diploma Projects
Below you can find out the work that is being undertaken by our current RMS Diploma students:
Elucidating the artifacts and abberations of SIM and STORM super resolution imaging technologies and subsequent protocol optimisation for cells and tissue sections
Julian Mark Bowen, University of Cambridge
Once our super resolution system is installed I will attempt to identify the reasons for artifacts and aberrations within the images and develop and optimise methods to eliminate them. Sample preparation is known to be critical to obtain good images using widefield and confocal imaging but is especially important for super resolution imaging.
The Characterisation of Speciality Fibres Utilising 3-D SEM Constructed Images
Michael Brookes, University of Leeds
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.
Analysis of the Subcellular Localisation and Kinetics of Survivin in Human Cells by Fluorescence and Correlative Fluorescence-EM Microscopy
Denise McLean, University of Nottingham
The principle aim is for me to develop fluorescence microscopy skills, some of which are compatible with the electron microscopy skills of which I am an expert, thus the CLEM technique will be of great value and others that involve laser technology and analysis of protein kinetics using advanced fluorescence imaging.
Comparative analysis of the Zeiss and IQ Deltavision TIRF acquisition systems and analysis algorithms to optimise A₁- and A₃- adenosine receptor analysis
Seema Rajani, University of Nottingham
Total Internal Reflection Fluorescence (TIRF) is a specialist imaging technique which produces an evanescent wave to excite fluorescence molecules. The technique significantly reduces background and so is ideal for the investigation of processes near the cell membrane. In this project I will be using TIRF to track the movement and clustering of A₁- and A₃- adenosine receptors at the cell membrane. I will be comparing two different TIRF microscope systems - Zeiss TIRF3 and IQ Deltavision for image acquisition and a range of propriety and open source analysis to optimize analyse membrane organization of A₁- and A₃- adenosine receptors.
The Study of Marek’s Disease Virus In Feather Follicle Epithelium
Jennifer Simpson, The Pirbright Institute
Marek's disease virus (MDV) is an alphaherpesvirus which causes a lymphoproliferative disease in chickens. Inflection occurs by inhalation of infected dander and following a complex life cycle involving infection of B and T cells, the virus is shed from the feather follicle. Vaccination is available and prevents clinical disease however it does not prevent the shedding of virus. The feather follicle is important in the transmission of MDV as the feather follicle epithelium (FFE) is the only region where fully enveloped and infectious virus particles are assembled. This study aims to identify cell tropism and describe viral morphogenesis in the FFE using immunofluorescence and transmission electron microscopy.
Comparative Analysis of Cellular Localisation of Viral Glycoproteins and Capsid Protein in Transfected Cells
Helen Todd, The Moredun Research Institute
A9.5 is a gene unique to the wildebeest-associated MCF virus, Alcelaphine herpesvirus-1. A Haemagglutinin (HA) epitope-tagged construct has been generated that can be transfected into HEK293T cells and has been found to express the A9.5 protein by Western blot. A9.5 encodes a predicted glycoprotein with a potential N-terminal signal sequence but the protein sequence has no similarity to known proteins. In order to define the cellular localization of A9.5 we will compare it with the known cell-surface viral antigen glycoprotein B (gB) and the major virus capsid protein (ORF25), by transfection of HEK293T cells and analysis by fluorescence microscopy.
Optimisation of microwave-assisted sample processing for biomedical research using the Leica EM AMW microwave tissue processor
Anna Pielach, University of Oxford
This project will explore automated microwave tissue processing of biological samples for routine and 3D electron microscopy using the Leica EM AMW. With only two such machines currently in the UK, including one used for diagnostic work, there is need for testing and optimization of protocols for a range of specimens. I will perform a detailed comparative study in the context of biomedical research at Sir William Dunn School of Pathology. Culture cells and tissues will be processed in the AMW in parallel with standard manual processing to fine-tune the protocols for reliable processing of diverse specimens.
Characterisation of the Migratory Properties of Additives used in Acrylic Coating on Polymer Film
Craig Holliday, Innovia Films
Polypropylene film is often coated with acrylic nano-particle formulations which contain migratory additives designed to modify the surface properties of the material. Whilst it is possible to quantify the levels using infra-red spectroscopy, this only provides a bulk assay.
It is hoped that a method to image the dried acrylic coating and map additive distributions on the dried surface can be developed to allow a study of the interaction of bulk levels and drying temperature on the migratory behaviour. This can then be correlated to physical and optical characteristics to optimise manufacturing conditions.
Examination of air-sensitive iron selenide superconducting materials via transmission electron-microscopy and related analytical techniques
Jennifer Holter, University of Oxford
Iron selenide compounds have been demonstrated as promising superconducting materials. Understanding and refining superconducting properties in these complex multi-phase materials relies upon insightful material characterization. Analytical election microscopy is a key tool for answering the key structural and compositional questions relating to phase separation. Progress thus far has been limited due to the highly reactive air sensitive nature of these compounds. This study involves designing new preparation protocols for such materials with the aim of yielding representative TEM and SEM specimens
First of all, the RMS Diploma was fun, I have nothing but good things to say about it and I learnt a lot of microscopy and biology. The project I undertook for the Diploma was a cell death project (how to measure necroptosis.) The RMS Diploma has helped me a lot, opening several doors for me. I was able to get my name on 5 publications because of it and gave a talk at the flowcytometryUK meeting this year. I also helped out with a workshop on cell death at the same meeting.
Since the RMS Diploma I have taken over the light microscopy facility at ICR and merged it with the flow cytometry facility. The facility is now called “Flow Cytometry and Light Microscopy Core Facility”. This increased responsibility was a direct consequence of doing the RMS Diploma in my opinion. As I am now running the microscopy facility I was able to attend the AQLM course in Woods Hole this year. It is a great 10 day course on microscopy which enhanced my knowledge of microscopy a lot. I am very grateful to the institute for supporting me with this course since it is a large cost to attend it, but I think finishing the Diploma showed the ICR that I am serious about learning microscopy and working in the field which led to me acquiring the funding. We are also investing a lot in microscopy this year at ICR which will grow the facility and give me a lot more responsibilities.
I would say that a lot of these new responsibilities and opportunities in my working life have been made possible because I completed the RMS Diploma.