Super resolution imaging of protein dynamics and functions in physiology and disease

Lead Research Organisation: University of Leeds
Department Name: Astbury Centre

Abstract

A major challenge in biomedical research is to be able to see inside a cell with enough detail to be able to understand how proteins interact with each other, and what goes wrong in disease. Typically, to see where a protein is inside a cell, a researcher will use a microscope that illuminates the cells with visible light together with dyes to label the proteins. This type of microscope can see detail to about 200nm (1/5000th of a millimetre), but cellular organelles are commonly at least 4 fold smaller than this, and proteins at least 10 fold smaller. Imaging these organelles and proteins with a normal light microscope does not reveal enough detail about the organisation of proteins and organelles within a cell, or help us understand the normal cellular functions of the proteins, and how they are affected by disease. To overcome this problem, several new types of microscopy have been recently invented to help see the detailed organisation of proteins inside cells. The goal of our research is to build a new microscope and develop new dyes that will help us achieve this, and solve important biomedical questions that are impossible to solve with our existing microscopes.

Technical Summary

Three main approaches to super-resolution imaging, available as either custom built instrumentation or commercially, are STED (Stimulated Emission Depletion, ~60nm resolution in XY), SIM (Structured Illumination Microscopy, ~100nm resolution in XY) and PALM/STORM (Photo-Activated Localisation Microscopy/Stochastic Optical Reconstruction Microscopy, ~20nm resolution in XY). We are currently building a new microscope (with inward investment from the University) capable of PALM/STORM.
In this new proposal, we plan to build an innovative M2SIM microscope, that will allow multiplanar 3D SIM imaging of live cells as fast imaging rates, to capture high resolution images of dynamic processes inside cells. This will complement the PALM/STORM system by providing high resolution in live cells, at high speeds that are difficult to achieve with PALM. In addition, to maximise the opportunity from our inward investment in PALM, and to serve the general needs of the imaging community, we plan to develop novel caged-dyes and small quantum dots with an increased blinking rate, and develop new approaches to label and tag proteins of interest. Together, these new approaches will allow a range of biomedical researchers at Leeds, working in key strategic areas (Cancer, Cardiovascular, Musculoskeletal, Neuroscience) to answer important biomedical questions that cannot currently be answered using existing technologies.

Planned Impact

Beneficiaries for this research will include the microscopy community, and the biomedical science/clinical community. The proposed developments in probes for super-resolution imaging, and the new M2SIM microscope will both be of interest to other researchers using these techniques. The development of both of these technologies is clearly of high interest to other 'imagers' in the UK, as evidenced by the recent Bio-imaging UK meeting at the Wellcome Trust. The participants at that meeting were all very enthusiastic about sharing expertise in these techniques. Through MP's role as chair of the Life Science Committee at the Royal Microscopy Society, she will facilitate workshops to share expertise to ensure that others do benefit. There is additional potential for these approaches being taken up by the commercial private sector, as evidence by our collaboration with Olympus. The biomedical community will benefit from being able to use the technology to answer questions currently impossible to answer with existing standard light microscopy approaches.
These developments have the potential to contribute to the nation's wealth and health, by the value added potential of novel microscopies and their potential for commercialisation, and by the solving of new questions to help us understand important biomedical problems, and devise new approaches to treat disease.
Staff working on the project will be trained in cutting edge techniques, and gain extra professional skills through participating in training others, presenting their work, running workshops, and taking part in other professional development opportunities.
Realistically, by the end of the 5 years, we expect to be running a cutting edge microscopy facility, open to researchers both in Leeds and elsewhere, that we expect will be a flagship for the UK research in this area.

Publications

10 25 50
 
Description Gravitropic setpoint angle control in higher plants
Amount £461,000 (GBP)
Funding ID BB/N010124/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 07/2016 
End 06/2019
 
Description Super-resolution imaging 
Organisation National Institute of Biomedical Imaging and Bioengineering
Country United States 
Sector Public 
PI Contribution We are collaborating with Hari Shroff to develop super-resolution imaging
Collaborator Contribution Hari Shroff is collaborating with us to develop super-resolution imaging
Impact Research paper in 'Methods' was published in 2015. Multidisciplinary (Hari Shroff - Engineering, Michelle Peckham, Biomedical/Biological)
Start Year 2014
 
Description Cafe Scientifique 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact I gave a talk on imaging to the York Philosophical Society, which sparked lots of questions and discussion.
Year(s) Of Engagement Activity 2016
URL http://www.thebasementyork.co.uk/#!yps-cafe-scientifique/c107b
 
Description Food: Truth or Scare 
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 Media (as a channel to the public)
Results and Impact I recorded an interview, with the BBC, about the subject of nutrition and hair as part of the program Food: Truth or Scare
Year(s) Of Engagement Activity 2019
URL https://www.bbc.co.uk/programmes/b08f17c0
 
Description In Our Time 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact I was an invited guest on 'In Our Time' on BBC Radio 4
Year(s) Of Engagement Activity 2013
URL http://www.bbc.co.uk/programmes/b03jdy3p
 
Description Invited Speaker at one day conference 'a history of the small', Oxford 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact The conference covered all aspects of 'the small' from microscopy to splitting the atom. I gave a talk on microscopy, and there was lively discussion and questions afterwards.
Year(s) Of Engagement Activity 2019
URL https://www.stx.ox.ac.uk/happ/events/history-small-one-day-conference
 
Description North West Science Network presentation at South Cheshire College 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact I gave a talk on Microscopy to the 6th form on microscopy
Year(s) Of Engagement Activity 2017
 
Description Royal Microscopical Society 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact I became the president of the Royal Microscopical Society
Year(s) Of Engagement Activity 2016
 
Description Royal Society MP Pairing Scheme 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Policymakers/politicians
Results and Impact I took part in the Royal Society MP Pairing Scheme in November 2015. I visited parliament, and shadowed an MP (Chris Green from Bolton West). Chris is due to make a return visit to the University in April 2016.
Year(s) Of Engagement Activity 2015,2016
URL https://royalsociety.org/grants-schemes-awards/pairing-scheme/
 
Description Soapbox, Art and Science 
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 This was a public event, as part of 'light night' in Leeds, where I teamed up with an artist (Jane Scott) to show up the use of microscopy in science and art, and the importance of women's contributions to research!
Year(s) Of Engagement Activity 2017
URL http://soapboxscience.org/soapbox-art-science-leeds-2017/
 
Description Super-resolution Imaging Workshops 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact We have run two super-resolution workshops at Leeds, as general discussion workshops for those scientists working in 'super-resolution' light microscopy. These workshops have resulted in much debate and discussion. The first workshop also led to a collection of articles being published in the journal 'Methods'.
Year(s) Of Engagement Activity 2014,2015
URL http://www.rms.org.uk/discover-engage/event-calendar/super-resolution-workshop-2015.html