MICA: Multi-parametric and super-resolution imaging of amyloidogenic proteins

Lead Research Organisation: University of Cambridge
Department Name: Chemical Engineering and Biotechnology

Abstract

Alzheimer's, like other neurodegenerative diseases, is caused by aberrant forms of proteins which tend to accumulate and aggregate into shapes, which are believed to be toxic to the brain cells of affected patients. The two proteins thought to cause Alzheimer's are called amyloid-beta and tau. The precise mechanisms by which these proteins aggregate into toxic forms are unknown, but there is accumulating evidence that the shape and size of forming aggregates is intricately linked to the onset and progression of disease. Yet there are no tools to study the growth of toxic aggregates in living cells, they are simply too small to be observed with traditional techniques. For the development of potential therapies against Alzheimer's, a microscopic understanding of the events leading to aggregation, and their consequence on affected cells, is crucial. Being able to observe protein aggregation in brain cells would permit us to relate the appearance of protein "clumps" with their toxic effect. Do affected cells malfunction? In what way? What aggregate species cause the cells to malfunction? Can we identify the toxic species leading cells to 'misbehave'? Can we inhibit the formation of "toxic shapes" in living cells through the addition of potential therapeutic agents? Can we stop the spreading of toxic species from cell-to-cell? Answers to these questions require radically new approaches to look into cells, at unprecedented resolution. In our group we have recently developed a new kind of microscope, a so called optical super-resolution microscope, that is so powerful that it is capable of visualising the shape of protein aggregates directly in cultured brain cells, which we use as model systems of disease. We were the first researchers to demonstrate that it is possible to obtain 'images' of toxic protein shapes in neurones, and here we propose to use this newly developed tool to shed new light on the mechanisms that lead to the onset and propagation of disease.
The aims of our proposed research project are: 1) to identify specific species of amyloid-beta and tau in neurones and other cells present in the brain and to relate the shape, and size of these species to the appearance of disease symptoms. 2) to study amyloid-beta and tau trafficking in and out of cells and to infer on the implications of this on the possible mechanisms of disease progression. 3) to see whether potential inhibitors of protein aggregation and propagation have an effect on what we observe within the cells we study. We will test several promising anti-aggregation anti-propagation compounds that our collaborators provide.
Through the development and application of advanced microscopic imaging techniques developed in our group we aim to gain insight into the molecular level processes that lie at the heart of Alzheimer's disease and other forms of dementia. Our techniques enable us to investigate the mechanisms that lead to the misfolding and spreading of proteins and their aggregation into toxic structures in live cell model systems at very high resolution, giving insight into the mechanisms of disease and its propagation. This understanding may help in the future search for potential therapeutic agents. The knowledge and tools generated here will provide a rational basis for the development of biomarkers/diagnostics that will enable physicians to: 1) detect the disease in its earliest stages when only biochemical changes are apparent; 2) identify the stage of the disease in order to tailor therapies for individual patients; and 3) monitor response to that treatment. Knowledge of these pathways will also provide targets for therapies to repair these pathways, thereby preventing or even reversing the disease.

Technical Summary

Amyloid proteins such as Abeta and tau are known to misfold and, eventually, to aggregate into insoluble deposits.

Questions that remain unsolved include: What are the neurotoxic amyloid species? Can the misfolded state propagate from one cell to another and has this an impact on the neuropathology? Do extracellular chaperones impact on degradation or aggregation pathways of amyloid species? The applicant's group specialises in the development and application of advanced microscopy techniques for the functional study of protein self-assembly reactions in neurodegenerative disease. We will use these tools to address the following questions:

- Abeta and tau aggregates are likely to be composed of ensembles of oligomers with different sizes and conformations and potentially differing neurotoxicity. We will use novel fluorescence-based sensors to: a) characterise the biophysical properties of these ensembles in live cells with a spatial resolution on the molecular scale; and b) correlate their biophysical properties with effects on neuropathology.

- There is increasing evidence in the literature that intracellular Abeta plays an important role in disease. In addition, the propagation of tau pathology appears to be crucial in spreading the disease to non-affected brain areas. We aim to characterise the trafficking mechanisms responsible for amyloid proteins being taken up by cells and/or released in the extracellular space.

- We will screen for different inhibitors of Abeta or tau mediated neuropathology. Firstly, we will investigate different inhibitors of amyloid proteins propagation. Secondly, we will test the potential of extracellular chaperones to interfere with amyloid protein degradation or aggregation.

Planned Impact

Over 820,000 people were affected by dementia in the UK in 2010, 24 million worldwide in 2005. This number is forecast to increase rapidly, making it one of the most pressing health issues in developed countries. The annual cost of dementia was over £23 billion in 2009. Our research is part of a global effort to assist the discovery of a cure for these devastating and costly diseases. Through the development and application of advanced microscopic imaging techniques developed in our group we aim to gain a mechanistic insight into the molecular level processes that lie at the heart of Alzheimer's disease and other forms of dementia. Our techniques enable us to investigate the mechanisms that lead to the misfolding of proteins and their aggregation into toxic structures in live cell model systems at molecular scale resolution, giving insight into the mechanisms of disease and its propagation. This understanding may help in the future search for potential therapeutic agents. The potential beneficiaries from this research are therefore:
- Neurobiologists studying neurodegenerative diseases (see previous section on academic beneficiaries)
- Pharmaceutical companies designing drugs will benefit from a better understanding of protein misfolding, aggregation, and dysfunction. They will also benefit from the technology we develop which could be optimised into methods for screening novel drugs and the investigation of their mode of action.
- Patients suffering from Alzheimer's disease and other forms of dementia, their carers and the community at large will benefit from a better understanding of the molecular and cellular events, which are involved in the onset and propagation of dementia. Drugs that may develop from such an understanding would greatly increase their quality and dignity of life as well as that of their relatives and carers.
- The project provides career opportunities for the named researchers through involvement and training in a cutting edge research enviroment on a topic of great socioeconomic importance. They will be trained in the development and application of state of the art molecular imaging methods, which finds use both in academic as well as in industrial research environments (e.g. in pharmaceutical companies).
- The National Physics laboratory will benefit through our collaboration in the development of optical super-resolution methods: the application of these techniques to address question of biomedical importance will provide incentives to further develop these methods (see letter of support). As the UK's national provider of measurement services, NPL will serve industry and academia on the whole with the provision of these technologies.
- The Neuroscience Consortium led by Prof. P. St. George-Hyslop, comprising research groups from across the UK and abroad will also benefit. Our technologies are critical to support the activities of these researchers, and our infrastructure and technical expertise will be made available to them free of charge.

Publications

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Blacker TS (2017) Investigating State Restriction in Fluorescent Protein FRET Using Time-Resolved Fluorescence and Anisotropy. in The journal of physical chemistry. C, Nanomaterials and interfaces

 
Description "THE ROLE OF THE ALZHEIMER'S DISEASE RISK GENE PICALM IN BLOOD BRAIN BARRIER TRANSPORT, ALZHEIMER'S DISEASE AND AMYLOID ANGIOPATHY "
Amount £760,940 (GBP)
Funding ID MR/N012453/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 04/2016 
End 05/2019
 
Description 2019 exercise
Amount £400 (GBP)
Organisation European Biophysical Societies' Association 
Sector Charity/Non Profit
Country Germany
Start 02/2018 
End 02/2018
 
Description Alzheimer Research UK Cambridge Network travel grant
Amount £500 (GBP)
Organisation Alzheimer's Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2013 
End 10/2013
 
Description British Biophysical Society Travel Grant
Amount £309 (GBP)
Organisation British Biophysical Society 
Start 05/2018 
End 05/2018
 
Description British Council Newton-Katip Celebi fund travel grant (Miranda Robbins)
Amount £200 (GBP)
Organisation British Council 
Sector Charity/Non Profit
Country United Kingdom
Start 04/2016 
End 04/2017
 
Description Cambridge ARUK Travel Grant
Amount £488 (GBP)
Organisation Alzheimer's Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 05/2018 
End 05/2018
 
Description Cambridge ARUK travel grant
Amount £300 (GBP)
Organisation Alzheimer's Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 03/2019 
End 03/2019
 
Description Cambridge ARUK travel grant (Miranda Robbins)
Amount £150 (GBP)
Organisation Alzheimer's Research UK 
Department Alzheimers Research UK, Cambridge
Sector Charity/Non Profit
Country United Kingdom
Start 03/2017 
End 03/2017
 
Description Cambridge Infinitus Research Centre
Amount £5,000,000 (GBP)
Organisation Infinitus 
Sector Private
Country China
Start 09/2015 
End 08/2020
 
Description EBSA bursary
Amount £330 (GBP)
Organisation European Biophysical Societies' Association 
Sector Charity/Non Profit
Country Germany
Start 02/2018 
End 02/2018
 
Description Liquid droplets and hydrogels in health and disease
Amount £400,000 (GBP)
Funding ID 203249/Z/16/Z 
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2017 
End 12/2021
 
Description Liquid droplets and hydrogels:protein phasetransition in health and disease
Amount £3,111,483 (GBP)
Funding ID 203249/Z/16/Z 
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 02/2017 
End 01/2021
 
Description Newnham College Travel Grant
Amount £731 (GBP)
Organisation University of Cambridge 
Department Newnham College
Sector Academic/University
Country United Kingdom
Start 06/2018 
End 09/2018
 
Description Oon Khye Beng Ch'Hia Tsio Studentships (Suil Collins)
Amount £1,500 (GBP)
Organisation University of Cambridge 
Department Downing College, Cambridge
Sector Academic/University
Country United Kingdom
Start 09/2016 
End 09/2017
 
Description Research Fellowship Deutsche Forschungsgemeinschaft (DFG)
Amount € 140,400 (EUR)
Funding ID LA 3609/2-1 
Organisation German Research Foundation 
Sector Public
Country Germany
Start 04/2015 
End 03/2018
 
Description SUPUVIR Marie Curie Consortium
Amount € 4,017,699 (EUR)
Funding ID 722380 
Organisation European Commission 
Sector Public
Country European Union (EU)
Start 01/2017 
End 12/2021
 
Description Swiss National Science Foundation
Amount SFr. 72,000 (CHF)
Organisation Swiss National Science Foundation 
Sector Public
Country Switzerland
Start 10/2014 
End 04/2016
 
Description UK-KOREA COLLABORATION FOR NEURODEGENERATIVE DISEASE
Amount £9,490 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 05/2016 
End 04/2017
 
Description Building of a dSTORM set up with Prof Marcus Sauer 
Organisation University of Wurzburg
Country Germany 
Sector Academic/University 
PI Contribution We contributed by building our own dSTORM set up and exchanging on the use of this technique for the study of neurodegenerative diseases
Collaborator Contribution Prof Marcus Sauer contributed by exchanging on his knowledge of the dSTORM technique and the photochemistry associated with it
Impact This collaboration resulted in our group having its own dSTORM set up Kaminski-Schierle GS, Sauer M, Kaminski CF, "Probing Amyloid Aggregation and Morphology In Situ by Multiparameter Imaging and Super-Resolution Fluorescence Microscopy," (2014) in Uversky VN, Lyubchenko YL, (eds) Bio-nanoimaging: Protein Misfolding & Aggregation, Academic Press, pp. 105-120 Kaminski Schierle GS, van de Linde S, Erdelyi M, Esbjörner EK, Klein T, Rees E, Bertoncini CW, Dobson CM, Sauer M, and Kaminski CF, "In Situ Measurements of the Formation and Morphology of Intracellular ß-Amyloid Fibrils by Super-Resolution Fluorescence Imaging", J. Am. Chem. Soc., 133 (33), pp 12902-12905, (2011).
Start Year 2008
 
Description Cambridge Imperial Centre for Single Cell Analysis (T. Knowles, F. Hollfelder, J. Edel) 
Organisation Imperial College London
Country United Kingdom 
Sector Academic/University 
PI Contribution We have written a joint grant proposal to set up a brain on chip platform.
Collaborator Contribution Joint grant proposal.
Impact Joint grant proposal written.
Start Year 2016
 
Description Cambridge Imperial Centre for Single Cell Analysis (T. Knowles, F. Hollfelder, J. Edel) 
Organisation University of Cambridge
Department MRC Cancer Unit
Country United Kingdom 
Sector Public 
PI Contribution We have written a joint grant proposal to set up a brain on chip platform.
Collaborator Contribution Joint grant proposal.
Impact Joint grant proposal written.
Start Year 2016
 
Description Computational study of the instrinsic fluorescence of amyloid fibrils with Ali Hassanali and Luca Grisanti 
Organisation University of Trieste
Country Italy 
Sector Academic/University 
PI Contribution Experimental study of the fluorescence properties of amyloid and peptide fibrils
Collaborator Contribution Ab initio molecular dynamics simulations of the modeled peptide fibril systems (computational study)
Impact Paper accepted in JACS http://dx.doi.org/10.1021/jacs.5b11012 (multidisciplinary: spectroscopy, optical physics and theoretical chemistry)
Start Year 2013
 
Description Graphene microelectrode arrays for simultaneous electrical and fluorescence measurements on neurons with Antonio Lombardo and Andrea Ferrari 
Organisation University of Cambridge
Department Cambridge Graphene Centre
Country United Kingdom 
Sector Academic/University 
PI Contribution Growing and maintaining neuronal cell cultures, and imaging them on the graphene devices
Collaborator Contribution Fabricating the graphene microelectrode arrays, and making electrical measurements on them
Impact Multidisciplinary activity between biotechnology and electrical engineering
Start Year 2014
 
Description Huntingtin propagation (Prof. G. Bates, Prof. H. Lashuel) 
Organisation King's College London
Country United Kingdom 
Sector Academic/University 
PI Contribution We are sharing a PDRA who is funded by the CHDI foundation. We are collaborating on producing recombinant Huntingtin in order to determine the seeding capacity of the protein, which we will studied by TIRF microscopy.
Collaborator Contribution Gillian Bates was awarded the founding for the PDRA by the CHDI foundation.
Impact The collaboration is multi disciplinary, involving optical microscopy and biology. Prof. Hilal Lashuel from EPFL in Switzerland has joined this collaboration as he is an expert in the production of recombinant Huntingtin.
Start Year 2015
 
Description Huntingtin propagation (Prof. G. Bates, Prof. H. Lashuel) 
Organisation Swiss Federal Institute of Technology in Lausanne (EPFL)
Country Switzerland 
Sector Public 
PI Contribution We are sharing a PDRA who is funded by the CHDI foundation. We are collaborating on producing recombinant Huntingtin in order to determine the seeding capacity of the protein, which we will studied by TIRF microscopy.
Collaborator Contribution Gillian Bates was awarded the founding for the PDRA by the CHDI foundation.
Impact The collaboration is multi disciplinary, involving optical microscopy and biology. Prof. Hilal Lashuel from EPFL in Switzerland has joined this collaboration as he is an expert in the production of recombinant Huntingtin.
Start Year 2015
 
Description Imaging of amyloid proteins present in cerebrospinal fluid with Prof Mathias Jucker 
Organisation German Centre for Neurodegenerative Diseases
Country Germany 
Sector Public 
PI Contribution We contributed by imaging the samples provided by the collaborators on our dSTORM optical nanoscopy microscope.
Collaborator Contribution The group of Prof Mathias Jucker contributed samples of cerebrospinal fluid from mice and from humans
Impact An original paper has been published and further collaboration will take place. This is a collaboration between a lab which has access to rodent and human cerebrospinal fluid (our collaborators) and our lab which develops optical nanoscopy equipment.
Start Year 2013
 
Description Integrated Microfluidic- Nanosensor System for Neuro- Functional Imaging and Neurotransmission Detection (Dr. E. Nugent) 
Organisation University of Cambridge
Department Department of Physics
Country United Kingdom 
Sector Academic/University 
PI Contribution Jane is currently setting up a brain on chip platform to study amyloid propagation in live neurons.
Collaborator Contribution Dr. Eileen Nugent has been co-supervising Jane.
Impact Eileen Nugent, Clemens F. Kaminski and Gabriele S. Kaminski Schierle: Super-resolution imaging of alpha-synuclein polymorphisms and their potential role in neurodegeneration, Integr. Biol., 2017, Advance Article, doi:10.1039/C6IB00206D
Start Year 2016
 
Description Interactions between aSynuclein and synaptic vesicles 
Organisation Swiss Federal Institute of Technology in Lausanne (EPFL)
Country Switzerland 
Sector Public 
PI Contribution Prepared recombinant protein and purified synaptic vesicles for interaction studies
Collaborator Contribution Microfluidics-integrated nanophotonics-enhanced IR spectroscopy platform
Impact Paper in preparation. Molecular/structural biology and bionanophotonics
Start Year 2018
 
Description Investigating the role of water in protein structure and aggregation 
Organisation Swiss Federal Institute of Technology in Lausanne (EPFL)
Country Switzerland 
Sector Public 
PI Contribution Prepared recombinant protein alpha synuclein for aggregation studied in the presence of different salts. Morphology studies
Collaborator Contribution Used second harmonic scattering to determine mobility of water around fibrils
Impact Paper in preparation. Molecular/structural biology and physics/biophotonics
Start Year 2018
 
Description Investigating the role of water in protein structure and aggregation 
Organisation University of Cambridge
Country United Kingdom 
Sector Academic/University 
PI Contribution Prepared recombinant protein alpha synuclein for aggregation studied in the presence of different salts. Morphology studies
Collaborator Contribution Runing and analysis of small angle neuron scattering data
Impact Paper in preparation, application for SANS beam time. Molecular biology and structural biology
Start Year 2018
 
Description Investigating the role of water in protein structure and aggregation 
Organisation University of Vermont
Country United States 
Sector Academic/University 
PI Contribution Prepared recombinant protein alpha synuclein for aggregation studied in the presence of different salts. Morphology studies
Collaborator Contribution Molecular dynamic simulations of peptide fragments in different salts
Impact Paper in preparation. Molecular/structural biology and physics/chemistry
Start Year 2018
 
Description Mechanism of Interaction of Synaptic Vesicles Induced by a synuclein (Dr. DeSimone) 
Organisation Imperial College London
Department Department of Life Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution Our experiments, using super-resolution microscopy, revealed an inherent molecular mechanism enabling aS to promote the interaction between synaptic vesicles.
Collaborator Contribution They examined the validity of this mechanism by rationally designing and experimentally testing a variant of aS that was engineered to promote stronger interactions between vesicles.
Impact Fusco G, Pape T, Stephens AD, Mahou P, Costa A, Kaminski CF, Kaminski Schierle GS, Vendruscolo M, Veglia G, Dobson CM, De Simone A, "Structural basis of synaptic vesicle assembly promoted by a-synuclein", Nature Communications. (2016), 7: 12563. doi:10.1038/ncomms12563 There is currently a manuscript written up for a follow up project.
Start Year 2015
 
Description Molecular chaperones as inhibitors of protein aggregation (Dr. Janine Kirstein, Berlin) 
Organisation Leibniz Association
Department Leibniz-Institute for Molecular Pharmacology
Country Germany 
Sector Public 
PI Contribution I have been invited to give a talk at the Leibniz-Institute for Molecular Pharmacology in Berlin
Collaborator Contribution A student from Dr. Kirstein's group will spend a Summer in my lab to test their chaperone related protein aggregation drugs
Impact not yet
Start Year 2016
 
Description SIM imaging of brain slices to investigate myelination with Robin Franklin 
Organisation Wellcome Trust
Department Wellcome - MRC Cambridge Stem Cell Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution Setting up the microscope, operation of the microscope, processing, reconstruction and analysis of the raw data, creation of a semi-automated image analysis protocol in Icy.
Collaborator Contribution Slicing of mouse brains, secondary anti-body staining, mounting of brain slices onto coverslip, statistical analysis of the data, biological interpretation.
Impact No publications yet, but promising results presented at various small talks - eg. in college, group meetings, to the IPES CDT
Start Year 2016
 
Description Secondary nucleation of monomers on fibril surface dominates a-synuclein aggregation and provides autocatalytic amyloid amplification (Prof Emma Sparr/Prof Sara Linse) 
Organisation Lund University
Department Department of Immunotechnology
Country Sweden 
Sector Hospitals 
PI Contribution Kinetic imaging of secondary nucleation reactions at the single molecule level with two colour direct stochastic Superresolution microscopy (dSTORM)
Collaborator Contribution Kinetic modelling of amyloid elongation reactions. Performed measurements in bulk samples.
Impact Paper in press at Quarterly Reviews of Biophysics Multidisciplinary: Chemistry, Physics
Start Year 2015
 
Description Setting up a brain on chip platform for the study of prion like propagation in amyloid-related diseases (Prof. Knowles) 
Organisation University of Cambridge
Department Department of Physics
Country United Kingdom 
Sector Academic/University 
PI Contribution Jane has been designing a new microfluidic based platform to sort and trap single neurons and to grow and differentiate them for live cell imaging. For this she has produced several PDMS devices, optimised flow conditions to guarantee fluidic isolation and is currently testing cell viability in these devices. She has further designed and implemented a device incubation chamber that can easily be adapted to fit on various microscope stages.
Collaborator Contribution Prof. Knowles PhD student Lianne Roode has been involved in this project and has helped with the design and evaluation of the original microfluidic design.
Impact We are currently writing up a manuscript on this.
Start Year 2016
 
Description Single molecule translation imaging with Christine Holt and Bill Harris 
Organisation University of Cambridge
Department Department of Physiology, Development and Neuroscience
Country United Kingdom 
Sector Academic/University 
PI Contribution Carried out microscopy imaging and provided tools for data analysis. Continuous feedback for improvement of the imaging and preparation conditions.
Collaborator Contribution Prepared samples of Xenopus eyes for the study of the development of neural tract in the frog. Provided the biological question.
Impact None. Ongoing collaborations
Start Year 2014
 
Description Structure of monomeric aSynuclein 
Organisation University of Antwerp
Country Belgium 
Sector Academic/University 
PI Contribution Prepared recombinant protein alpha synuclein for aggregation studied in the presence of different salts. Morphology studies
Collaborator Contribution native mass spec and ion mobility mass spec
Impact Two papers in preparation. Molecular biology and structural biology
Start Year 2018
 
Description Structure of monomeric aSynuclein 
Organisation University of Exeter
Country United Kingdom 
Sector Academic/University 
PI Contribution Prepared recombinant protein and purified synaptic vesicles for interaction studies
Collaborator Contribution Hydrogen-Deuterium Exchange Mass Spectroscopy
Impact Paper in preparation. Molecular biology and structural biology
Start Year 2016
 
Description Structure of monomeric aSynuclein 
Organisation University of Leeds
Country United Kingdom 
Sector Academic/University 
PI Contribution Prepared recombinant protein alpha synuclein for aggregation studied in the presence of different salts. Morphology studies
Collaborator Contribution native mass spec and ion mobility mass spec
Impact Two papers in preparation. Molecular biology and structural biology
Start Year 2018
 
Description Study of monomeric Tau propagation from cell to cell with Prof Eckard Mandelkow 
Organisation German Centre for Neurodegenerative Diseases
Country Germany 
Sector Public 
PI Contribution We have applied multi parametric and super resolution imaging techniques to the study of Tau propagation from cell to cell
Collaborator Contribution Prof Eckard Mandelkow provided us with purified recombinant Tau protein, both unlabelled and fluorescently labelled.
Impact Michel CH, Kumar S, Pinotsi D, Tunnacliffe A, St George-Hyslop P, Mandelkow E, Mandelkow E-M, Kaminski CF, Kaminski Schierle GS, "Extracellular Monomeric Tau is Sufficient to Initiate the Spread of Tau Pathology", J. Biol. Chem. (2014), 289: 956-967. This collaboration is multi-disciplinary and involved biochemistry and physics
Start Year 2010
 
Description Super-resolution imaging of macromolecules micelles assemblies with Ian Manners 
Organisation University of Bristol
Department School of Chemistry
Country United Kingdom 
Sector Academic/University 
PI Contribution Carried out super-resolution microscopy (dSTORM) imaging and feedbacks on how to prepare and design fluorescent probes for preparation of their micellar samples.
Collaborator Contribution Designed, prepared and synthetised the micellar structure used for the study of micelle elongation.
Impact None yet. But manuscript under preparation.
Start Year 2014
 
Description Super-resolution imaging of nanotubes involved in the transfer of Tau from neuron to neuron (Luc Buee) 
Organisation National Institute of Health and Medical Research (INSERM)
Department Lille (INSERM)
Country France 
Sector Public 
PI Contribution We would carry out super-resolution imaging on samples
Collaborator Contribution Will provide cells, plasmids for expression of Tau and expertise in nanotubes
Impact Invited talk given by Dr Gabriele Kaminski Schierle in Lille for a seminar on New technologies and Neurosciences
Start Year 2015
 
Description Synthetic 3D Neuronal Networks (Prof. Heutnik, Dr. Cesare) 
Organisation Eberhard Karls University of Tubingen
Department Natural and Medical Sciences Institute
Country Germany 
Sector Academic/University 
PI Contribution We have written a proposal that we submitted to the VW Foundation.
Collaborator Contribution Joint proposal to VW Foundation.
Impact Joint proposal submitted
Start Year 2016
 
Description Synthetic 3D Neuronal Networks (Prof. Heutnik, Dr. Cesare) 
Organisation German Centre for Neurodegenerative Diseases
Country Germany 
Sector Public 
PI Contribution We have written a proposal that we submitted to the VW Foundation.
Collaborator Contribution Joint proposal to VW Foundation.
Impact Joint proposal submitted
Start Year 2016
 
Description Synthetic 3D networks 
Organisation German Centre for Neurodegenerative Diseases
Department Tübingen Research Campus
Country Germany 
Sector Private 
PI Contribution Written joint proposal to VW Foundation.
Collaborator Contribution Written joint proposal to VW Foundation.
Impact Written joint proposal to VW Foundation.
Start Year 2016
 
Description Terrahertz spectroscopy of amyloid fibrils (Prof. Axel Zeitler) 
Organisation University of Cambridge
Country United Kingdom 
Sector Academic/University 
PI Contribution We have formed L-Glu based nanostructures which display fluorescent properties.
Collaborator Contribution L-Glu based nanowires are currently investigated by THz spectroscopy in order to determine the origin of the fluorescence.
Impact First data sets are currently analysed.
Start Year 2017
 
Description The Identification and development of small molecule inhibitors for the aggregation of amyloid beta - David Spring 
Organisation University of Cambridge
Department Department of Chemistry
Country United Kingdom 
Sector Academic/University 
PI Contribution The project was developed by a joint PhD researcher within the three groups. Project planning, microscopy and cell work are carried out in our group.
Collaborator Contribution - Project planning and the development of microfluidic techniques are carried out in the Hollfelder group. - The Spring group contributed small molecule compounds for screening.
Impact - Output: A joint PhD student who carries out the majority of the experimental work between the three laboratories. The project is multi-diciplinary. Microscopy and cell work are carried out in our group. Microfluidic assay development is being carried out in the the Hollfelder group. Organic synthesis of the small molecule screening libraries was carried out in the Spring group - Outcomes: None yet, but the research is still actively providing results which are expected to be submitted for publishing within the next few months.
Start Year 2014
 
Description Translational cell biology of new microglial-related dementia genes (P. St. George Hyslop, O. Paulsen, G. Mallucci, D. Rubinsztein, D. Klenerman) 
Organisation University of Cambridge
Country United Kingdom 
Sector Academic/University 
PI Contribution We have developed a microfluidic based neuronal platform in which we can amyloid protein propagation. We are currently setting up a new microfluidic device which will permit patch-clamp recordings to be taken of two individually (within the microfluidic device) connected neurons. This device will permit super-resolution recordings of propagating amyloid proteins to be imaged simultaneously with the electrophysiological recording.
Collaborator Contribution Patch clamp recordings will be undertaken by the collaborating partner.
Impact Written a joint grant application.
Start Year 2017
 
Description Why do biological nanowires glow? (Dr. D. Credgington) 
Organisation University of Cambridge
Department Department of Physiology, Development and Neuroscience
Country United Kingdom 
Sector Academic/University 
PI Contribution We applied for a nanoCDT studentship.
Collaborator Contribution Application for nanoCDT studentship to study the intrinsic fluorescence related to amyloid proteins.
Impact Application for nanoCDT studentship.
Start Year 2017
 
Description aSynuclein-lipid interaction 
Organisation University of Cambridge
Country United Kingdom 
Sector Academic/University 
PI Contribution Prepared recombinant alpha synuclein and brain extract lipid samples for aSyn/membrane interactions studies. Performed AFM analysis.
Collaborator Contribution Access to FastScan AFM in Pharmacology
Impact Paper in preparation. Molecular biology and structural biology
Start Year 2018
 
Description dSTORM imaging of DNA origami with Ulrich Keuser 
Organisation University of Cambridge
Department Cavendish Laboratory
Country United Kingdom 
Sector Academic/University 
PI Contribution Carried out super-resolution microscopy imaging using DNA origami nano-structure. Carried out analysis and simulations based on observation proving the feasibility of the imaging.
Collaborator Contribution Designed, prepared and synthetised the DNA origami structure.
Impact None. Ongoing collaborations. Proof of principle of imaging of their DNA origami structures under our microscope. Foster further collaboration leading to offering student projects.
Start Year 2014
 
Description super-resolution imaging of amyloid proteins with Prof Sara Linse 
Organisation Lund University
Country Sweden 
Sector Academic/University 
PI Contribution The Laser Analytics Group is in the process of imaging amyloid protein fibril formation with super-resolution microscopy
Collaborator Contribution The group of Prof Sara Linse provided purified amyloid protein and gave a two-week tutorial to three members of the Laser Analytics group on purification of proteins
Impact This partnership is still young and we hope it will develop into publication on the amyloid fibril formation kinetics. This collaboration is multi-disciplinary, the Laser Analytics group being specialised in super-resolution imaging (physics, engineering) and the group or Prof Sara Linse in protein purification and biophysical characterisation of amyloid proteins
Start Year 2014
 
Description ARUK Spring appeal 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact We participated in the preparation of an appeal letter and follow up letter to potential funders of ARUK regarding the use of emergency funding by ARUK. This was also followed by a blog post

We expect some persons who received this appeal send fund to Alzheimer's Research UK for research on neurodegenerative diseases.
Year(s) Of Engagement Activity 2014
URL http://www.dementiablog.org/every-project-counts/
 
Description Blog post on ARUK website on dSTORM (2014) 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Engage with patients, carers and families about research funded by ARUK

Readers of the Alzheimer's Research UK might donate towards research.
Year(s) Of Engagement Activity 2014
URL http://www.dementiablog.org/10000-times-smaller-pinhead/
 
Description CamBRAIN : Teaching children about neurons 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Teaching children about neurons
Year(s) Of Engagement Activity 2018
 
Description Episode of Elemental Ideas on Cambridge TV 25 Minute TV show about super-resolution microscopy 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Clemens Kaminski, Gabriele Kaminski Schierle, Colin Hockings, Florian Ströhl, Nathan Curry

Episode of Elemental Ideas on Cambridge TV 25 Minute TV show about super-resolution microscopy
01/07/2016 Explaining research to the public through a video of our group's lab and research work, with interviews of the two group leaders and several postdoctoral research associates and PhD students.
The videos can be accessed for free online.

Elemental Ideas is Cambridge TV's science magazine programme. They've produced more than 50 programmes since August 2015, covering research topics including string theory, diabetes, exoplanets, vulcanology, epigenetics and the wildlife of New Zealand.
Year(s) Of Engagement Activity 2016
URL http://www.cambridge-tv.co.uk/super-resolution-microscopy/
 
Description Interview by ARUK at the Copenhagen AAIC conference 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact This activity resulted into engaging with readers of the Alzheimer's Research UK blog about research presented by Dr Claire Michel on a poster at the Alzheimer's Association International Conference 2014

With this interview funders of Alzheimer's Research UK received information regarding how their donations are used by researchers and what results researchers obtain.
Year(s) Of Engagement Activity 2014
 
Description Interview on BBC Radio Cambridge 2014 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact This activity was a 3 minutes interview on super-resolution interview used in research on neurodegenerative diseases

Engage with the wide public about advances in research on Alzheimer's disease
Year(s) Of Engagement Activity 2014
 
Description Pint of Science Festival Cambridge 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Public lecture at Pint of Science Festival.
Year(s) Of Engagement Activity 2016
 
Description PreLighter 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Writing highlight sections on interesting preprints for the PreLight website and twitter feed
Year(s) Of Engagement Activity 2018,2019
 
Description Research Photography Exhibition 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Finalist in STEM photography competition at Hughes Hall College, Cambridge
Year(s) Of Engagement Activity 2019
 
Description Research exchange meeting with DAAD students 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Undergraduate students
Results and Impact Organised a visit (lab tours and research lectures) for German Biophysics students funded under the "Studienstiftung" programme
Year(s) Of Engagement Activity 2016
 
Description STEM in Song 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact Encouraging school-aged girls to engage with the STEM subjects through music. Awards event with science demos, song launch at the LMB, music video at https://www.youtube.com/watch?v=7c4KMOWoZW4 . Reach: 15300 (according to Facebook insights)
Year(s) Of Engagement Activity 2018
URL https://www.youtube.com/watch?v=7c4KMOWoZW4
 
Description Seminar on Science and Brewing 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Explaining science to the public
Year(s) Of Engagement Activity 2018
 
Description Showcasing research to public in a hands on manner at the Cambridge Science Festival 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Cambridge Science Festival
Showcasing research to public in a hands on manner through various experiments organised in the dept of Chemistry including looking at neurons under a microscope.
Explaining the group's research to the public
Undertaken by several members of the group on a yearly basis since 2016

The Science Festival provides the public with opportunities to explore and discuss issues of scientific interest and concern and to raise aspirations by encouraging young people to consider a career in science, technology, engineering or mathematics.

Each year, the Festival welcomes visitors to hundreds of events and receives extensive national and local media coverage. Over 170 event coordinators organise talks, interactive demonstrations, hands-on activities, film showings and debates with the assistance of around 1,000 staff and students from departments and organisations across the University and research institutions, charities and industry in the eastern region. In addition, over 150 people volunteer their time to act as stewards to ensure visitors have a safe and enjoyable Festival experience.
Year(s) Of Engagement Activity 2016,2017
URL http://www.sciencefestival.cam.ac.uk/
 
Description So you want to be a scientist?' experiment and speaking presentation 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Talking to school children about life as a scientist and GCSE options to take
Year(s) Of Engagement Activity 2018
 
Description Superresolution public video 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Video interview and research video to explain superresolution microscopy in medical research to a lay audience. Currently >14000 views.
Year(s) Of Engagement Activity 2015
URL https://www.youtube.com/watch?v=W-0GWbOFT3w
 
Description Talk and lab tour to funders of ARUK, Feb 2016 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Supporters
Results and Impact Engage with funders on the research carried out thanks to their donations
Year(s) Of Engagement Activity 2016
 
Description Talk and lab tour to funders of ARUK, May 2015 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Supporters
Results and Impact Engage with funders on the research carried out thanks to their donations
Year(s) Of Engagement Activity 2015
 
Description Talk on A short history of Microscopy, Cambridge 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Public lecture on A short history of Microscopy
Year(s) Of Engagement Activity 2017
 
Description Talk, tour of the lab and Q&A for 15 employees of ARUK 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact This activity gave a clearer understanding to employees of ARUK and to readers of the ARUK blog of the work carried out by researchers studying dementia

One employee of ARUK wrote a blog post on his visit, which increased the number of people reached through this activity.
Year(s) Of Engagement Activity 2014
URL http://www.dementiablog.org/tau-proteins/
 
Description Tech Me Out team volunteer 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Pint of Science Festival Cambridge
Year(s) Of Engagement Activity 2018