Advanced mass spectrometry imaging to identify the biological mechanisms underlying the functions of surface lipids in Drosophila

Lead Research Organisation: University College London
Department Name: MRC Laboratory of Molecular Cell Biology

Abstract

The skin of animals, from humans to Drosophila, is coated with a complex layer of lipids that have a range of functions including minimising trans-epithelial water loss and also providing a barrier against microbial infection. It is not yet clear which lipid species are anti-microbial and which prevent dehydration. Recent research in our lab has also identified a novel function of these lipids in Drosophila; certain classes have been shown to be autotoxic, however the biological mechanism underlying this is not yet fully understood. These questions represent a significant metrological challenge, and to answer these we will use the 3D OrbiSIMS. This novel imaging mass spectrometry instrument combines high lateral resolution and mass accuracy to provide detailed spatial information about the chemical composition of the sample which is not possible using existing techniques. The development of ambient and cryogenic methodology for biological samples using this instrument will provide great progress to the field of biological imaging. In particular, the cryo-SIMS methodology developed will be critical for the accurate measurement of biological samples providing a state as close to native as possible to reduce experimental artefacts.

The aims and objectives of this project are to:
Develop 3D OrbiSIMS imaging methodology for lipids and metabolites on the Drosophila cuticle. This involves the development of cryogenic sample preparation methods.
Develop data analysis pipelines for identification of lipids and small molecules using the high mass resolving power and mass accuracy, as well as MS/MS capacity for increased assignment confidence.
Identify the function of different lipid species, and the biological mechanism via which certain species can cause toxicity.

Imaging mass spectrometry methodology will be developed using the 3D OrbiSIMS at the National Physical Laboratory. This instrument integrates the high lateral resolution of ToF-SIMS and the high mass resolving power and mass accuracy of an Orbitrap analyser with MS/MS capabilities, alongside 3D depth-profiling and image reconstruction to create a 3D chemical image of the sample. We will use this technique, combined with the power of Drosophila genetics, to answer key questions on the role of cuticular lipids.

This research aligns with the EPSRC's Healthcare Technologies strategy to develop cross-cutting research capabilities, in particular the development of novel imaging techniques. The development of 3D OrbiSIMS lipid and metabolite imaging will provide unparalleled 3D chemical imaging of biological samples with combined high lateral resolution and mass resolving power. The use of this cutting-edge technique to investigate the distribution and function of Drosophila cuticular lipids fits well into the Analytical Science research area of the EPSRC which is interested in the development of novel techniques to analyse biological matter and systems.

This research is a collaboration between the Francis Crick Institute providing the biological expertise, and the National Physical Laboratory providing the expertise in imaging mass spectrometry.

Publications

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Studentship Projects

Project Reference Relationship Related To Start End Student Name
EP/P510683/1 01/10/2016 30/09/2021
1938905 Studentship EP/P510683/1 26/09/2016 30/09/2020 Clare Newell
 
Description General Travel Grant
Amount £303 (GBP)
Organisation Biochemical Society 
Sector Learned Society
Country United Kingdom
Start 09/2018 
End 09/2018
 
Description Rowland Hill Award
Amount € 500 (EUR)
Organisation Ionoptika 
Sector Private
Country United Kingdom
Start 09/2017 
End 09/2017
 
Description National Physical Laboratory - National Centre of Excellence in Mass Spectrometry Imaging 
Organisation National Physical Laboratory
Country United Kingdom 
Sector Academic/University 
PI Contribution Clare is a PhD student based part time at the National Physical Laboratory. She contributes intellectually to the project and contributes her experience in the application of mass spectrometry imaging to biological tissues gained through her joint placement at the National Physical Laboratory and the Francis Crick Institute.
Collaborator Contribution The PhD is secondarily supervised at the National Physical Laboratory by Ian Gilmore. The Gilmore lab contribute their significant knowledge of mass spectrometry imaging and intellectually contribute to this project. The lab also contribute access to mass spectrometry imaging instrumentation including the 3D OrbiSIMS. The lab have provided training for Clare on various mass spectrometry imaging techniques.
Impact This collaboration has enabled participation in the following outreach activities: NPL Open House and the Royal Society Summer Science Exhibition. The intellectual and instrumentational contribution from this collaboration has resulted in the award of Young Surface Analyst of the Year 2018. This is a multi-disciplinary collaboration contributing knowledge in physics and chemistry.
Start Year 2016
 
Description The Francis Crick Institute 
Organisation Francis Crick Institute
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution Clare is a PhD student based at the Francis Crick Institute. She contributes intellectually to the project and contributes her experience in the application of mass spectrometry imaging to biological tissues gained through her joint placement at the National Physical Laboratory and the Francis Crick Institute.
Collaborator Contribution The PhD is primarily supervised at the Francis Crick Institute by Alex Gould. Alex and other members of the Gould lab contribute intellectually to this project, especially through their significant experience using Drosophila melanogaster as a model organism. The lab and the Francis Crick Institute contribute access to equipment and consumables. The Metabolomics Science and Technology Platform at the Francis Crick Institute contribute their expertise in mass spectrometry and access to various instruments located at the Crick.
Impact This collaboration has enabled participation in the following outreach activities: New Scientist Live and Science Museum "Cellulates". The intellectual contribution from this collaboration has resulted in the award of Young Surface Analyst of the Year 2018. This is a multi-disciplinary collaboration contributing knowledge in biology.
Start Year 2016
 
Description NPL Open House 
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 Research institute was open to the general public for half a day, including the imaging mass spectrometry laboratory. Members of the public were able to learn about the new instrumentation that is currently being developed, and the applications we use it for. School students also attended the day and this increased interest in studying STEM subjects including Biology and Physics.
Year(s) Of Engagement Activity 2017
URL http://www.npl.co.uk/open-house/
 
Description New Scientist Live 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Participated in running a stand aimed at educating the general public about CRISPR gene editing technology. The stand increased public understanding of the method and helped to tackle many ethical questions surrounding the use of the technique. Many members of the public came away with a better understanding and changed their opinions on the ethics of the technique due to uncovering pre-existing misconceptions.
Year(s) Of Engagement Activity 2017
URL https://live.newscientist.com/new-scientist-live-2017?page=6
 
Description Royal Society Summer Science Exhibition 
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 Participated in running a stand which aimed to communicate the concept of imaging mass spectrometry to the general public. This sparked many discussions including possible applications, and increased interest in the area especially among school students. This increased the general understanding of many students, most whom had not been exposed to such techniques before. It also helped increase public understanding of the importance of fundamental research, and how this could be applied in the future.
Year(s) Of Engagement Activity 2017
URL https://royalsociety.org/science-events-and-lectures/2017/summer-science-exhibition/exhibits/
 
Description Science Museum "Cellulates" 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Participated in running a stall educating the general public about the applications of fruitflies and fruitfly genetics in current research. The aim was to increase the visibility of this experimental model and it increased public understanding of the relevance of this model and how we can use flies to reduce the number of mammals needed in research. Many members of the general public came away with an increased understanding of fruitfly genetics, and many changed their opinions on the relevance of this model for the better.
Year(s) Of Engagement Activity 2017
URL https://www.sciencemuseum.org.uk/see-and-do/lates
 
Description St George's University of London Careers Panel 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Undergraduate students
Results and Impact 50 students and alumni attended a panel discussion where sandwich placements and careers in STEM were discussed. The audience were composed of biomedical science undergraduate students and through attending the panel discussion, many solidified their plans to persue a career in academia.
Year(s) Of Engagement Activity 2019
 
Description St George's University of London Placement Programmes 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Undergraduate students
Results and Impact 50 students attended a series of talks about the merits of undertaking a sandwich year at a research institution in a laboratory and the benefits this may have to their future careers. As a result, several students were encouraged to apply to the Crick sandwich year programme.
Year(s) Of Engagement Activity 2018