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 two question 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. 

Andrew Jarjour image STED scaled down

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. 

Sean BrownCalcium 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.

DeeFigure 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.

Miguel crop3D 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.

Judithβ-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.

Tushar cropConfocal 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.

Combined StacksGranules (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.

STED MBFigure 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:

Date Journal Title DOI Author List
2021/03 Journal of Cell Biology Parameter-free molecular super-structures quantification in single-molecule localization microscopy 10.1083/jcb.202010003 M. Marenda, E. Lazarova, S. van de Linde, N. Gilbert & D. Michieletto
2020/11 Nature Genetics cGAS-mediated induction of type I interferon due to inborn errors of histone pre-mRNA processing 10.1038/s41588-020-00737-3 C. Uggenti, A. Lepelley, M. Depp, A.P. Badrock, M.P. Rodero, M. El-Daher, G.I. Rice, S. Dhir, A.P. Wheeler, A. Dhir, W. Albawardi, M. Frémond, L. Seabra, J. Doig, N. Blair, M. Martin-Niclos, E. Della Mina, A. Rubio-Roldán, J.L. García-Pérez, D. Sproul, J. Rehwinkel, J. Hertzog, A. Boland-Auge, R. Olaso, J. Deleuze, J. Baruteau, K. Brochard, J. Buckley, V. Cavallera, C. Cereda, L..M.H. De Waele, A. Dobbie, D. Doummar, F. Elmslie, M. Koch-Hogrebe, R. Kumar, K. Lamb, J.H. Livingston, A. Majumdar, C.M. Lorenço, S. Orcesi, S. Peudenier, K. Rostasy, C.A. Salmon, C. Scott, D. Tonduti, G. Touati, M. Valente, H. van der Linden Jr., H. Van Esch, M. Vermelle, K. Webb, A.P. Jackson, M.A. M. Reijns, N. Gilbert & Y.J. Crow
2020/08 bioRxiv PrePrint SuperStructure: a Parameter-Free Super-Structure Identifier for SMLM Data 10.1101/2020.08.19.254540 M. Marenda, E. Lazarova, S. van de Linde, N. Gilbert & D. Michieletto
2020/07 Developmental Cell PRL3-DDX21 Transcriptional Control of Endolysosomal Genes Restricts Melanocyte Stem Cell Differentiation 10.1016/j.devcel.2020.06.013 J.A. Johansson, K. L. Marie, Y. Lu, A. Brombin, C. Santoriello, Z. Zeng, J. Zich, P. Gautier, A. von Kriegsheim, H. Brunsdon, A.P. Wheeler, M. Dreger, D.R. Houston, C.M. Dooley, A.H. Sims, E.M. Busch-Nentwich, L.I. Zon, R.S.Illingworth & E.E. Patton
2020/03 Soft Matter Chromosome compaction and chromatin stiffness enhance diffusive loop extrusion by slip-link proteins 10.1039/C9SM01875A A. Bonato, C.A. Brackley, J. Johnson,  D. Michieletto and  D. Marenduzzo
2019/11 Molecular Cell Decreased Enhancer-Promoter Proximity Accompanying Enhancer Activation

 

1016/j.molcel.2019.07.038

 

N.S. Benabdallah, I. Williamson, R.S. Illingworth, L. Kane, S. Boyle, D. Sengupta, G.R. Grimes, P. Therizols & W.A. Bickmore
2019/01 The Journal of Biological Chemistry A VPS33A-binding motif on syntaxin17 controls autophagy completion in mammalian cells

1074/jbc.RA118.005947

R.S Saleeb, D.M. Kavanagh, A.R.Dun, P.A.Dalgarno, R.R. Duncan
2018/09 Scientific Reports Super-silent FRET Sensor Enables Live Cell Imaging and Flow Cytometric Stratification of Intracellular Serine Protease Activity in Neutrophils

1038/s41598-018-31391-9

T.H. Craven, N. Avlonitis, N. McDonald, T. Walton, E. Scholefield, A.R. Akram, T. S. Walsh, C. Haslett, M. Bradley & K. Dhaliwal
2018/09 The EMBO Journal Mitochondrial inner membrane permeabilisation enables mtDNA release during apoptosis

15252/embj.201899238

J.S. Riley, G. Quarato, C. Cloix, J. Lopez, J. O'Prey, M. Pearson, J. Chapman, H. Sesaki, L.M. Carlin, J.F. Passos, A.P. Wheeler, A. Oberst, K.M. Ryan, S.W.G. Tait
2018/09 Translational Psychiatry DISC1 regulates N-methyl-D-aspartate receptor dynamics: abnormalities induced by a Disc1 mutation modelling a translocation linked to major mental illness 10.1038/s41398-018-0228-1 E.L.V. Malavasi, K.D. Economides, E. Grünewald, P. Makedonopoulou, P. Gautier, S. Mackie, L.C. Murphy, H. Murdoch, D. Crummie, F. Ogawa, D.L. McCartney, S.T. O’Sullivan, K. Burr, H.S. Torrance, J. Phillips, M. Bonneau, S.M. Anderson, P. Perry, M. Pearson, C. Constantinides, H. Davidson-Smith, M. Kabiri, B. Duff, M. Johnstone, H.G. Polites, S.M. Lawrie, D.H. Blackwood, C.A. Semple, K.L. Evans, M. Didier, S. Chandran, A.M. McIntosh, D.J. Price, M.D. Houslay, D.J. Porteous & J.K. Millar
2018/01 The EMBO Journal Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy

15252/embj.201695709

K.N. Richter, N.H. Revelo, K.J. Seitz, M.S. Helm, D. Sarkar, R.S. Saleeb, E. D'Este, J. Eberle, E. Wagner, C. Vogl, D.F. Lazaro, F. Richter, J. Coy‐Vergara, G. Coceano, E.S. Boyden, R.R. Duncan, S.W. Hell, M.A. Lauterbach, S.E. Lehnart, T. Moser, T.F. Outeiro, P. Rehling, B. Schwappach, I. Testa, B. Zapiec, and S.O. Rizzoli
2017/12 IEEE Transactions on Electron Devices A 256 x 256 100-kfps 61% Fill-Factor SPAD Image Sensor for Time-Resolved Microscopy Applications doi:10.1109/TED.2017.2779790 I. Gyongy, N. Calder, A. Davies, N. Dutton, R.R. Duncan, C. Rickman, P. Dalgarno, R.K. Henderson
2017/12 Scientific Reports A Bi-fluorescence complementation system to detect associations between the Endoplasmic reticulum and mitochondria

1038/s41598-017-17278-1

M. Harmon, P. Larkman, G. Hardingham, M. Jackson & P. Skehel
2017/10 Scientific Reports High-throughput assessment of mechanical properties of stem cell derived red blood cells toward cellular downstream processing

1038/s41598-017-14958-w

E. Guzniczak, M.M. Zadeh, F. Dempsey, M. Jimenez, H. Bock, G. Whyte, N. Willoughby & H. Bridle
2017/06 Cell SAF-A Regulates Interphase Chromosome Structure through Oligomerization with Chromatin-Associated RNAs

1016/j.cell.2017.05.029

R. Nozawa, L. Boteva, D.C. Soares, C. Naughton, A.R. Dun, A. Buckle, B. Ramsahoye, P.C. Bruton, R.S. Saleeb, M. Arnedo, B. Hill, R.R. Duncan, S.K. Maciver & N. Gilbert
2017/03 eLife Ambra1 spatially regulates Src activity and Src/FAK-mediated cancer cell invasion via trafficking networks

7554/eLife.23172

C. Schoenherr, A. Byron, E. Sandilands, K. Paliashvili, G.S. Baillie, A. Garcia-Munoz, C. Valacca, F. Cecconi, B. Serrels & M.C. Frame
2017/01 Renal Physiology - American Journal of Physiology Bimodal dynamics of granular organelles in primary renin-expressing cells revealed using TIRFmicroscopy

 

1152/ajprenal.00384.2016

 

A.R. Dun, G.J. Lord, R.S. Wilson, D.M. Kavanagh, K.I. Cialowivz, S. Sugita, S. Park, L. Yang, A.M. Smyth, A. Papadopulos, C. Rickman & R.R. Duncan
2017/01 Scientific Reports Tryptophan and Non-Tryptophan Fluorescence of the Eye Lens Proteins Provides Diagnostics of Cataract at the Molecular Level

1038/srep40375

A. Gakamsky, R.R. Duncan, N.M. Howarth, B. Dhillon, K.K. Buttenschön, D.J. Daly & D. Gakamsky
2017/01 Nature Communications Repo-Man/PP1 regulates heterochromatin formation in interphase

1038/ncomms14048

I.J. de Castro, J. Budzak, M.L. Di Giacinto, L. Ligammari, E. Gokhan, C. Spanos, D. Moralli, C. Richardson, J.I. de las Heras, S. Salatino, E.C. Schirmer, K.S. Juri Rappsilber, C.G. Ullman, W.A. Bickmore, S. Lamble, M.W. Goldberg, V. Vinciotti & P. Vagnarelli
2016/11 Open Biology SBE6: a novel long-range enhancer involved in driving sonic hedgehog expression in neuralprogenitor cells

1098/rsob.160197

N.S. Benabdallah, P. Gautier, B. Hekiimoglu-Balkan, L.A. Lettice, S. Bhatia & W.A. Bickmore
2016/11 Scientific Reports Smart-aggregation imaging for single molecule localisation with SPAD cameras

1038/srep37349

I. Gyongy, A. Davies, N.A. Dutton, R.R. Duncan, C. Rickman, R.K. Henderson & P.A. Dalgarno
2016/10 Genes and Development Condensin II mutation causes T-cell lymphoma through tissue-specific genome instability

1101/gad.284562.116

J. Woodward, G.C. Taylor, D.C. Soares, S. Boyle, D. Sie, D. Read, K. Chathoth, M. Vukovic, N. Tarrats, D. Jamieson, K.J. Campbell, K. Blyth, J.C. Acosta, B. Yistra, M.J. Arends, K.R. Kranc, A.P. Jackson, W.A. Bickmore & A.J. Wood
2016/08 Development Shh and ZRS enhancer co-localisation is specific to the zone of polarizing activity

1038/ncomms3056

H. Patel, J.Zich, B. Serrels, C. Rickman, K.G. Hardwick, M.C. Frame & V.G. Brunton

2016/05 American Journal of Human Genetics A Restricted Repertoire of De Novo Mutations in ITPR1 Cause Gillespie Syndrome with Evidence for Dominant-Negative Effect

1016/j.ajhg.2016.03.018

M. McEntagart, K.A. Williamson, J.K. Rainger, A.P. Wheeler, A. Seawright, E. De Baere, H. Verdin, L.T. Bergendahl, A. Quigley, J. Rainger, A. Dixit, A. Sarkar, E. López Laso, R. Sanchez-Carpintero, J. Barrio, P. Bitoun, T. Prescott, R. Riise, S. McKee, J. Cook, L. McKie, B. Ceulemans, F. Meire, I.K. Temple, F. Prieur, J. Williams, P. Clouston, A.H. Németh, S. Banka, H. Bengani, M. Handley, E. Freyer, A. Ross, DDD Study, V. van Heyningen, J.A. Marsh, F. Elmslie & D.R. FitzPatrick
2016/04 Scientific Reports PSD95 nanoclusters are postsynaptic building blocks in hippocampus circuits

1038/srep24626

M.J. Broadhead, M.H. Horrocks, F. Zhu, L. Muresan, R. Benavides-Piccione, J. DeFelipe, D. Fricker, M.V. Kopanitsa, R.R. Duncan, D. Klenerman, N.H. Komiyama, S.F. Lee & S.G.N. Grant
2016/01 Scientific Reports Translation Microscopy (TRAM) for super-resolution imaging

 

1038/srep19993

 

Z. Qiu, R.S. Wilson, Y. Liu, A.R. Dun, R.S. Saleeb, D. Liu, C. Rickman, M.C. Frame, R.R. Duncan & W. Lu
2015/12 Biofabrication Three-dimensional bioprinting of complex cell laden alginate hydrogel structures.

 

1088/1758-5090/7/4/045012

 

A. Ghanizadeh Tabriz, M.A. Hermida, N. Leslie & W. Shu
2015/12 Cell Reports ADF and Cofilin1 Control Actin Stress Fibers Nuclear Integrity and Cell Survival. 1016/j.celrep.2015.10.056 G. Kanellos, J. Zhou, H. Patel, R.A. Ridgway, D. Huels, C.B. Gurniak, E. Sandilands, N.O. Carragher, O.J. Sansom, W. Witke, V.G. Brunton & M.C. Frame
2015/11 IEEE Transactions on Biomedical Engineering Estimation of Fluorescence Lifetimes via Rotational Invariance Techniques

1109/TBME.2015.2491364

C Buckley, AR Dun, A Peter, C Bellamy, KW Gross, RR Duncan & JJ Mullins
2015/11 Scientific Reports SIMcheck: a Toolbox for Successful Super-resolution Structured Illumination Microscopy

1038/srep15915

H. Yu, R. Saleeb, P. Dalgarno & D. Day-Uei Li
2015/09 eLife TALPID3 controls centrosome and cell polarity and the human ortholog KIAA0586 is mutated in Joubert syndrome (JBTS23)

7554/eLife.08077

L.A. Stephen, H. Tawamie, G.M. Davis, L. Tebbe, P. Nürnberg, G. Nürnberg, H. Thiele, M. Thoenes, E. Boltshauser, S. Uebe, O. Rompel, A. Reis, A.B. Ekici, L. McTeir, A. Fraser, E. Hall, P. Mill, N. Daudet, C. Cross, U. Wolfrum, A.R. Jamra, M.G. Davey & H.J. Bolz
2015/09 Genes & Development A negative loop within the nuclear pore complex controls global chromatin organization

1101/gad.264341.115

 
M. Breuer & H. Ohkura
2015/08 Histochemistry and Cell Biology Chromatin at the nuclear periphery and the regulation of genome functions

1007/s00418-015-1346-y

C. Lemaître & W.A. Bickmore
2015/06 Methods The evolution of structured illumination microscopy in studies of HIV

1016/j.ymeth.2015.06.007

K. Marno, L. Al’Zoubi, M. Pearson, M. Posch, Á. McKnight & A. P. Wheeler
2015/03 Angewandte Chemie Rapid formation of a supramolecular polypeptide-DNA hydrogel for in situ three-dimensional multilayer bioprinting.

1002/anie.201411383

C. Li, A. Faulkner-Jones, A. R. Dun, J. Jin, P. Chen, Y. Xing, Z. Yang, Z. Li, W. Shu, D. Liu & R.R. Duncan
2014/12 Nature Communications A molecular toggle after exocytosis sequesters the presynaptic syntaxin1a molecules involved in prior vesicle fusion.

1038/ncomms6774

D.M. Kavanagh, A.M. Smyth, K.J. Martin, A. Dun, E.R. Brown, S. Gordon, K.J. Smillie, L.H. Chamberlain, R.S. Wilson, L. Yang, W. Lu, M.A. Cousin, C. Rickman & R.R. Duncan
2014/10 Journal of Cell Science Eps8 controls Src- and FAK-dependent phenotypes in squamous carcinoma cells. 10.1242/jcs.157560 C. Schoenherr, B. Serrels, C. Proby, D.L. Cunningham, J.E. Findlay, G.S. Baillie, J.K. Heath & M.C. Frame
2013/09 Frontiers in Endocrinology Imaging large cohorts of single ion channels and their activity

3389/fendo.2013.00114

K. Hiersemenzel, E.R. Brown & R.R. Duncan
2013/08 Science Transcription. Flashing a light on the spatial organization of transcription

10.1126/science.1242889

C. Rickman & W.A. Bickmore
2013/02 The Journal of Biological Chemistry Munc18-1 protein molecules move between membrane molecular depots distinct from vesicle docking sites

1074/jbc.M112.407585

A.M. Smyth, L. Yang, K.J. Martin, C. Hamilton, W. Lu, M.A. Cousin, C. Rickman & R.R. Duncan
2013/01 Nature Communications Kindlin-1 regulates mitotic spindle formation by interacting with integrins and Plk-1

1038/ncomms3056

H. Patel, J. Zich, B. Serrels, C. Rickman, K.G. Hardwick, M.C. Frame & V.G. Brunton