ESRIC welcomes researchers from a range of disciplines and institutes, below are some examples of a few of these projects as well as the list of publications that have come out of the facility since 2013. We love to hear how you are getting on and you can fill us in at this quick 2 questions survey here or enter any new publications at your ppms account page here.

Polarity signalling in central nervous system myelin

Professor Charles ffrench-Constant 
Dr. Andrew Jarjour (PDRA)
Centre for Regenerative Medicine, University of Edinburgh

This project used the multicolour and 3-Dimensional super-resolution technique, STED, to visualise polarity signalling in oligodendroglia at the regions flanking the nodes of ranvier. Proper polarization of the myelin sheath at these paranodal regions is essential for axo-glial adhesion and the resultant organization of the axon into the distinct molecular domains required for rapid impulse conduction. Understanding the mechanisms involved in polarizing the myelin sheath requires that the myelin sheath be imaged in fine detail, which as seen in the image, STED can provide. This work has significant influence on our understanding of how CNS myelin forms and the demyelination that is seen in patients with multiple sclerosis. 

Image of a nerve cell (red) with myelin wrapped around in a spiral (green) shows the level of detail in STED (left) compared to standard imaging (right)

Ion channels in sperm cells and their association with impaired fertility

Dr Sean Brown
School of Science, Engineering and Technology, Abertay University

This project aims to investigate the expression and intracellular localisation of ion channels inside human sperm as it has been found that abnormal channel phenotypes are associated with impaired fertilisation. In particular the knocking out of certain genes that relate to impaired fertility can drastically change calcium channel arrangement along the tail. This project is using single molecule localisation microscopy techniques to quantify the molecular organisation on the nanometer scale over 3 dimensions. 

Calcium channel localisation inside a human sperm cell imaged on the ESRIC Olympus TIRF system, right hand image shows zoom.

Cellular communication and diabetes

Dr Colin Rickman
Dr. Deirdre Kavanagh (PDRA)
Institute of Biological Chemistry, Bioengineering and Biophysics, Heriot-Watt University
Funded by MRC and Wellcome Trust

Using single molecule localisation microscopy such as PALM and STORM this project aims to understand the plasma membrane organisation of tSNARE proteins underlying insulin secretion in β-cells under normal physiological conditions and how this is altered in Type 2 related hyperglycemia. Previous work combined these techniques with Fluorescence Cross Correlation Spectroscopy to study SNARE protein interactions in neuronal cells.

Figure panel from: A molecular toggle after exocytosis sequesters the presynaptic ​syntaxin1a molecules involved in prior vesicle fusion, Nature Communications 2014.

3D Printing of Tumours

Dr Nick Leslie (PI)
Miguel Hermida (PhD)
Institute of Biological Chemistry, Bioengineering and Biophysics, Heriot-Watt University
Funded by Anonymous Trust

This project involves the development of a 3D printing strategy, which uses common biocompatible hydrogels to create multilineage tumour-like structures. These are visualised in ESRIC using live-cell 3D confocal imaging with the aim to study the tumour microenvironment and provide novel highly-reproducible models for preclinical drug testing.

3D printed construct in RGD-Alginate showing U87MG-EYFP glioblastoma cells, WI-38 Cerulean fibroblasts and activated MM-6 mCherry macrophages, 24h after printing. Imaged using the SP5 Confocal laser scanning microscope.

Distribution of pathogen associated molecular patterns on the Candida albicans cell surface

Prof. A. Brown (PI)
Dr Judith Bain (PDRA)
School of Medicine, Medical Sciences and Nutrition, University of Aberdeen
Funded by MRC

This project investigates the cell surface arrangement of pathogen associated molecular patterns (PAMPS), including β-glucan, on the wall of the fungal pathogen, Candida albicans. In this collaboration, Structured Illumination Microscopy (SIM) is used to visualise alterations in PAMP exposure in response to altered host factors.

β-glucan on the C. albicans cell surface, visualised by Fc-Dectin-1 using the SIM system

Analysis of human lung tissue and bacterial labelling compounds

Dr Kev Dhaliwal (PI)
Dr Tushar Choudhary (PDRA) 
Centre for Inflammation Research, University of Edinburgh
Funded by EPSRC

This collaborative project between Heriot-Watt University and Edinburgh University investigates the properties of markers for lung diseases, which forms part of the Proteus project; developing a tool for next-generation diagnostic testing of lung disease. Fluorescence Lifetime Imaging Microscopy (FLIM) is used to study the fluorescent properties of these markers.

Confocal images of fluorescently-labelled bacteria and the corresponding image showing the Fluorescence Lifetimes of each bacteria in a false colour scale

Investigating granular dynamics in renin-producing primary cells

Dr John Mullins (PI)
Dr Charlotte Buckley (PDRA) 
BHF Centre for Cardiovascular Science, University of Edinburgh
Funded by British Heart Foundation

The transport and dynamics of renin-producing granules inside primary juxtaglomerular cells has been investigated in a collaborative project between Universities. This research has been possible using TIRF microscopy and photoactivation techniques at ESRIC allowing live-cell tracking and quantification of calcium dynamics.

Granules (left panel) inside a renin-producing (right panel, green) juxtaglomerular cell imaged in live-cell conditions using the Ollympus IX81 TIRF system.

Super-Resolution imaging of postsynaptic proteins in the brain

Professor Seth Grant (PI - previous) and Dr Gareth Miles
Dr Matthew Broadhead (PDRA),
Centre for Neurodegeneration, University of Edinburgh and School of Psychology and Neuroscience, University of St. Andrews
Funded by Principals Career Development Scholarship

Gated stimulated emission depletion (g-STED) microscopy was used in this project to quantify PSD95eGFP organisation at the nanoscale in brain tissue to understand the underlying molecular framework of cognitive functions at the level of the synapse. This work was published in Nature Scientific Reports in May 2016 as part of a Wellcome Trust funded project at the University of Edinburgh. Matthew now works as a PDRA in St Andrews and continnues to collaborate with ESRIC.

Figure legend from above publication showing dye-filled neurons with single and multiple neuronal clusters in spine heads. Left panel, PSD95-eGFP (green) imaged with a confocal (Con) microscope and the Alexa Fluor 594-filled spine is shown in magenta. Right panel, g-STED image of PSD95-eGFP in same spine. Scale bars 500 nm.

ESRIC Publications: