3D OrbiSIMS: Label free chemical imaging of materials, cells and tissues

Lead Research Organisation: University of Nottingham
Department Name: Sch of Pharmacy

Abstract

This application is for a time of flight secondary ion mass spectrometer (ToF-SIMS) with unique state-of-the-art 3D imaging capability exhibiting unprecedented mass resolution achieved through the integration of a high specification OrbitrapTM mass spectrometer. Additionally, the instrument is capable of extremely high spatial resolution and is complemented by cryo-preparation facilities which allow the preservation of the native structure of hydrated samples such as biological cells and tissue. The instrument provides a label free molecular characterisation of materials using surface mass spectrometry of liberated secondary ion fragments generated by primary ion impaction from the outermost 1 - 2 nm. When this surface sensitivity is combined with a sputtering beam it produces a 3D chemical analysis of materials at high lateral (< 100 nm) and vertical (~ 3 nm) resolution.

The emerging next generation of real world systems and devices exhibit an increasing complexity in sample type throughout a variety of research areas, such as biomedical implants, drug delivery systems, organic electronics devices and engineering devices. The design and innovation of these devices is underpinned by materials characterisation, however their chemical complexity can be prohibitive to their characterisation. The instrument will offer an uncompromisingly accurate portrayal of the true chemical 3D internal environment of a given sample, specialising in the analysis of organic materials.

The detailed chemical characterisation of real world systems will have applications in a multi-disciplinary range of new research whilst supporting existing research programmes led by the PI and Co-Is within the Schools of Pharmacy, Life Sciences and Faculty of Engineering working in the areas of drug delivery, antimicrobial resistance and electronics amongst others. The chemically rich information in the ToF-SIMS experiment has been found to provide critical information in the performance of a range of real world material systems. The instrument operates under ultra-high vacuum and can be used to characterise solid samples of any given chemistry. Critically this is a label free approach, providing a full characterisation of the chemistry, unbiased by sample preparation choices and artefacts introduced by fluorophores employed in cell and tissue imaging by optical microscopy. Using the cryo -preparation facilities, the instrument will be world leading in its capability to analyse frozen hydrated liquids or semi-solids (for example, stem cells and bacteria) thereby ensuring that it can be used to analyse a very wide range of materials and is therefore truly transdisciplinary in its capacity.

The University of Nottingham is uniquely situated to house such an instrument with an international reputation in the application of ToF-SIMS in the pharmaceutical and materials sciences since the late 80s and active cryo-sample electron microscopy programmes which can be applied to maximise the utility of this combination. The University of Nottingham hosts a centralised facility where the instrument will be located with equipment access and importantly expertise provided for internal and external academic research. Internal academic research programmes that will be facilitated by this instrument including EPSRC Centres for Doctoral Training in Advanced Therapeutics & Nanomedicines, Carbon Capture and Storage and Cleaner Fossil Energy, Sustainable Chemistry, Additive Manufacturing and 3D Printing and Regenerative Medicine. Additionally, existing collaborative links will be exploited within the MI universities and amongst other national institutes to enable a step change in the 3D materials characterisation in areas such as pharmaceutics (Prof. Alistair Florence, University of Strathclyde), regenerative medicine (Prof. Molly Stevens, Imperial College London), semiconductor materials, devices and technology (Prof. David Wood, University of Durham).

Planned Impact

Economic Impact

The proposed facility will have a number of direct and indirect impacts upon various sectors of the UK economy. Direct impacts will arise from the use of the proposed facility to conduct contract research by such companies such as Walgreens Boots Alliance and Innospec Ltd to advance their research and development programmes in the healthcare and engineering sector respectively. These advances will be linked to new products and associated additional revenue generation. The facility will also be made available to consultancy companies, such as Juniper Pharmaceutical Services and Aystorm Ltd, who represent a large number of national and international clients in the pharmaceutical and high tech materials sectors respectively. The analytical work performed by such companies will generate direct revenue for the UK economy whilst maintaining the UK's strong reputation in this field.

Indirect impact will be delivered in the form of an enhanced capability for advanced research and development for industry in collaboration with Universities, including Nottingham, and others within the MI group and nationally including The University of Leeds, Durham, Manchester, Imperial, Sheffield and Glasgow. The scope of industrial collaboration is extensive and will utilise existing links to EPSRC funded PhD training programmes and EPSRC Centres for Innovative Manufacturing including those in Advanced Therapeutics and Nanomedicines, Regenerative Medicine and Sustainable Chemistry. These include >80 companies such as AZ, GSK, Innospec, Unilever, Rolls Royce, Dyson Appliance Ltd and Smith and Nephew. Relationships are being built with existing UK Catapult centres, e.g. Chris Herbert of the Cell Therapy Catapult, and will be developed with the Formulation Centre, for which the capabilities of the facility will be applicable to a wide range of industrial research and development needs.

Impact upon Society

The multi-disciplinary spread of the users of the proposed materials characterisation facility impacts many sectors of research, particularly within applied disciplines and as such will impact society in a variety of ways. The capability of the facility to enable research within the fields of pharmacy, tissue engineering and biomaterials will lead to the development of more effective drugs, controlled delivery systems, advanced biomaterials, and regenerative medicine technologies. These will produce improvements in the health and wellbeing of both UK and international society.

Enabling research in areas such as life sciences and pharmacy and engineering, the proposed facility will facilitate research into the development of antimicrobial resistance and antimicrobial resistant materials simultaneously in a true multi-disciplinary approach. Utilising current academic links to industry internationally the impact of the research can be realised and implanted both within the UK and more widely. Further materials science and physics related research areas including those within the field of graphene and semi-conductor research will also be enabled by the proposed ToF-SIMS facility ensuring that the UK remains competitive in state-of-the-art device manufacturing.

In all of the disciplines where the proposed facility will be active, the use of the instrument and the associated data processing will form a significant portion of the training and development of the next generation of research scientists. The use of the instrument in undergraduate projects and University held open days for local secondary schools will provide research experience and inspiration to the younger generation of potential scientists within the UK.
 
Description Whilst the grant is still in progress, some key findings have already been established.

The most notable of those is that the facility has been used to allow identification of proteins at solid surfaces. These include biomaterials and tissues sections. This is significant because this method allows the localisation of proteins in 3 dimensions. [Kotowska et al. Nature Communications 2020] Exploitation of this capability is now being investigated in bacterial biofilm studies, basic biological studies of matrix bound vesicles and the effects of drugs on local metabolism.

Most recently procedures used in metabolomics have been adopted to allow semi automated assignment of the majority of species present its he data rich OrbiSIMS spectra. This has recently been published in Edney et al Analytical Chemistry 2022. It is anticipated that this will become a routine tool for those using this technique.
Exploitation Route It is anticipated that the protein identification method will be used widely now published.

A list of collaborators are reflected in the papers published to date, ranging from archaeology, internal combustion engine additives, regenerative medicine, biology and medical device development.
Sectors Chemicals,Energy,Healthcare,Manufacturing, including Industrial Biotechology,Culture, Heritage, Museums and Collections,Pharmaceuticals and Medical Biotechnology,Transport

URL https://www.nottingham.ac.uk/research/groups/biomaterials-discovery/news/2020/nature-communications-2020.aspx
 
Description Research goes to new depths in skin peptide study https://www.nottingham.ac.uk/news/research-goes-to-new-depths-in-skin-peptide-study Tuesday, 25 May 2021 Researchers have revealed for the first time that a peptide in a leading beauty product can penetrate the tough stratum corneum barrier of the skin. A study between the University of Nottingham and No7, the UK's number one skincare brand, used leading-edge technology to track the penetration of No7's peptide blend Matrixyl 3000+™ following topical application to the skin surface. The findings offer an improved understanding of the bioavailability of topically applied peptides and provides further evidence for the efficacy of wrinkle targeting peptides. Researchers used a tape-stripping method to remove layers of skin from study participants, in vivo, whilst maintaining the integrity of layers and structural features. A recently developed hybrid secondary ion mass spectrometry technique, 3D OrbiSIMS, was employed in the blinded study and detected the peptide penetrating into the tough stratum corneum barrier - the major obstacle and rate limiting step for the delivery of active ingredients into the skin (n=12; p<0.05 vs untreated). The study used a recently popularised analytical technique to identify the peptide ten surface layers deep into the skin, whilst preserving the skin structure. Many other techniques rely on preparing skin samples for analysis by freeze-drying, fixing or staining, and therefore cannot be truly sure of the depth measured. "Never before have we been able to analyse the real-life penetration of the peptides in No7 in such detail," writes Dr Mike Bell, No7 skincare scientific advisor: For the first time, it has been possible to trace and detect the peptides responsible for tackling invisible photo-damage that occurs early in the ageing process. The 3D OrbiSIMS technique has opened up the possibility to track exactly where our peptides go to in the skin; now there are new insights into how to optimise delivery to make peptides even more effective. David Scurr, Senior Research Fellow, School of Pharmacy The flagship brand of the newly formed No7 Beauty Company, No7 was the first beauty brand in the UK to be able to demonstrate long term clinical and cumulative reductions in the appearance of lines and wrinkles on its Protect & Perfect Intense Advanced Serum. Since their creation, more than 5,000 women have been involved in the testing of all of No7's serums across nearly 80 trials. In scientifically controlled clinical trials, over 90% achieved a visible reduction in the appearance of wrinkles. The company's comprehensive approach to testing has proven that the serums dramatically slow the appearance of key signs of ageing, with advanced formulations that target damage; and now, its most recent testing with the 3D OrbiSIMS technique has given even greater insights into the powerful technology at the heart of the serums. The University of Nottingham was the first University in the world to own and operate the 3D OrbiSIMS instrument. The technology facilitates an unprecedented level of molecular analysis for a range of materials including biological tissues, such as human skin. Importantly, its high mass resolving power, chemical specificity and high sensitivity allow it to be used on human skin samples to identify and analyse low concentrations of active ingredients that resemble native components of the skin, such as collagen peptides. "This level of analysis of the Matrixyl 3000+™ peptide blend has not been possible until now. In the future, the technology may also allow us to identify new biochemical biomarkers of ageing and disease," says Dr Mike Bell. Story credits More information is available from Dr David Scurr on David.Scurr@nottingham.ac.uk
First Year Of Impact 2021
Sector Chemicals,Healthcare,Retail
Impact Types Societal,Economic

 
Description A New Correlative Approach for Structure Determination & Imaging of Molecular Materials
Amount £1,470,902 (GBP)
Funding ID EP/W006413/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 12/2021 
End 11/2026
 
Description BBSRC National Biofilms Innovation Centre Innovative Knowledge Centre
Amount £15,600,000 (GBP)
Funding ID BB/R012415/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2017 
End 09/2022
 
Description Designing bio-instructive materials for translation ready medical devices
Amount £5,200,000 (GBP)
Funding ID EP/X001156/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 01/2023 
End 12/2026
 
Description Dialling up performance for on demand manufacturing
Amount £7,200,000 (GBP)
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 10/2022 
End 09/2027
 
Description Dynamic physicochemical nanoscale imaging at the solid-liquid interface
Amount £1,171,682 (GBP)
Funding ID EP/V053884/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 08/2021 
End 07/2026
 
Description Enabling Next Generation Additive Manufacturing
Amount £5,852,466 (GBP)
Funding ID EP/P031684/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 10/2017 
End 09/2022
 
Description Enabling Next Generation Additive Manufacturing for Pharma and related industries
Amount £5,900,000 (GBP)
Funding ID EP/P031684/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 09/2017 
End 08/2022
 
Description High resolution, cryogenic analytical and transfer scanning electron microscope (HR-CAT-SEM)
Amount £1,564,542 (GBP)
Funding ID EP/S021434/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 04/2019 
End 03/2024
 
Description How simple plastic surfaces can be recruited to the fight against contact transmission of SARS-CoV-2
Amount £417,480 (GBP)
Funding ID EP/V055372/1 
Organisation United Kingdom Research and Innovation 
Sector Public
Country United Kingdom
Start 01/2021 
End 07/2022
 
Description Live monitoring of foreign-body response in animals by diffuse Raman spectroscopy
Amount £506,474 (GBP)
Funding ID NC/W001179/1 
Organisation National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) 
Sector Public
Country United Kingdom
Start 09/2021 
End 03/2024
 
Description NanoPrime: Maximising Equipment and Expertise Sharing in Nanoscience
Amount £202,437 (GBP)
Funding ID EP/R025282/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 04/2018 
End 07/2021
 
Description Targeting cannabinoids to the intestinal lymphatic system to improve treatment of multiple sclerosis and other autoimmune diseases
Amount £53,116 (GBP)
Funding ID M902 
Organisation Rosetrees Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 11/2019 
End 10/2021
 
Title A new particle mounting method for surface analysis Adam A. Dundas, Stefanie Kern, Valentina Cuzzucoli Crucitti, David J. Scurr, Ricky Wildman, Derek J. Irvine, Morgan R. Alexander First published: 08 September 2021 https://doi.org/10.1002/sia.7010 
Description Abstract The chemical analysis of microparticles is challenging due to the need to mount the particles on a substrate for analysis; double-sided adhesive tape is often used (sometimes conductive), however that is usually coated with poly (dimethyl siloxane) (PDMS) that is often used as a release agent. PDMS is a common surface contamination that can mask surface chemistries and hinder material performance where it is dependent on this contaminated interface. It is known that PDMS contains a very mobile oligomeric fraction that readily diffuses across surfaces resulting in the contamination of mounted particulate samples before and during surface chemistry analysis. This makes it impossible to determine whether the PDMS has arisen from the analysis procedure or from the sample itself. A new sample preparation method is proposed where polymer microparticles are mounted on a poly (hydroxyethyl methacrylate) (pHEMA) polymer solution, which we compare with particles that have been mounted on adhesive discs using time-of-flight secondary ion mass spectrometry (ToF-SIMS) and 3D OrbiSIMS analysis. Particles mounted on the pHEMA substrate results in a reduction of PDMS signal by 99.8% compared with microparticles mounted on adhesive discs. This illustrates how a simple, quick and inexpensive polymer solution can be used to adhere particles for analysis by ToF-SIMS, or other surface chemical analysis techniques such as X-ray photoelectron spectroscopy (XPS), without introduction of large amounts of silicone contaminant. 
Type Of Material Technology assay or reagent 
Year Produced 2022 
Provided To Others? Yes  
Impact It is hoped that this will be useful to people analysing the surface chemistry of particles. 
URL https://doi.org/10.1002/sia.7010
 
Title Cryo-OrbiSIMS for 3D molecular imaging of a bacterial biofilm in its native state 
Description We developed a method for analysis and imaging of biological samples in their native state, by combining a cryo-OrbiSIMS instrument with cryogenic sample handling and high-pressure freezing. By using this method, we did analysis and imaging of frozen-hydrated mature Pseudomonas aeruginosa biofilm, which allows the identification and map of quorum sensing signaling molecules, nucleobases and bacterial membrane molecules with high spatial-resolution and high mass-resolution. Some of quorum sensing signaling molecules were further confirmed by MS/MS. By comparing the analysis of frozen-hydrated Pseudomonas aeruginosa biofilm with the freeze-dried one, we dicover that signal intensity of all interesting molecules get enhanced in the frozen-hydrated state. Especially for polar molecules, such as amino acid, it could even achieve 10,000 fold increasing. Here, we provide the original OrbiSIMS data including MS and MS/MS spectra, depth profile and images of frozen-hydrated and freeze-dried Pseudomonas aeruginosa biofilm. The data could be open by using SurfaceLab Version 7.0 (ION-TOF, Germany). 
Type Of Material Database/Collection of data 
Year Produced 2020 
Provided To Others? Yes  
URL https://zenodo.org/record/3831954
 
Title Cryo-OrbiSIMS for 3D molecular imaging of a bacterial biofilm in its native state 
Description We developed a method for analysis and imaging of biological samples in their native state, by combining a cryo-OrbiSIMS instrument with cryogenic sample handling and high-pressure freezing. By using this method, we did analysis and imaging of frozen-hydrated mature Pseudomonas aeruginosa biofilm, which allows the identification and map of quorum sensing signaling molecules, nucleobases and bacterial membrane molecules with high spatial-resolution and high mass-resolution. Some of quorum sensing signaling molecules were further confirmed by MS/MS. By comparing the analysis of frozen-hydrated Pseudomonas aeruginosa biofilm with the freeze-dried one, we dicover that signal intensity of all interesting molecules get enhanced in the frozen-hydrated state. Especially for polar molecules, such as amino acid, it could even achieve 10,000 fold increasing. Here, we provide the original OrbiSIMS data including MS and MS/MS spectra, depth profile and images of frozen-hydrated and freeze-dried Pseudomonas aeruginosa biofilm. The data could be open by using SurfaceLab Version 7.0 (ION-TOF, Germany). 
Type Of Material Database/Collection of data 
Year Produced 2020 
Provided To Others? Yes  
URL https://zenodo.org/record/3831955
 
Description No. 7 Beauty Company (Walgreens Boots Alliance PLC) 
Organisation Boots UK
Country United Kingdom 
Sector Private 
PI Contribution A commercial study undertaken to assess the skin permeation of a cosmetic agent from a product produced by No. 7 Beauty Company. Analysis of human tape stripped samples using the 3D OrbiSIMS instrument to detect the delivery of a chemical agent.
Collaborator Contribution Provision of the human tape strip samples for analysis.
Impact Publication - Starr et. al., PNAS (2022) https://www.pnas.org/doi/10.1073/pnas.2114380119 Data used towards successful claims made by No. 7 Beauty Company and incorporated into Advertising Standards Agency approved media advertising. Telegraph article https://www.telegraph.co.uk/health-fitness/body/scientific-breakthrough-proves-anti-ageing-serums-work/ Reported in 13 media outlets https://pnas.altmetric.com/details/124934713/news
Start Year 2021
 
Description Facility opening event on 14/1/19 at the nmRC with invited national and international speakers 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Researchers from the UK but also one invited international speaker came to this launch event where the capabilities of this new facility was highlighted along with access arrangements and laboratory visits.
Year(s) Of Engagement Activity 2019
URL https://www.nottingham.ac.uk/isac/news-items/university-launches-3d-orbisims-at-nmrc.aspx
 
Description Fourth Annual Biomaterials Discovery Workshop 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact ~over 100 people attended the Third Biomaterials Discovery Workshop at the University of Nottingham in January 2020. The audience was mainly academic but also PhDs, Post Docs, Industry members etc. The BDW involved some excellent discussions in this area and will continue to be an annual event.
Year(s) Of Engagement Activity 2017,2018,2019,2020
URL https://www.nottingham.ac.uk/research/groups/biomaterials-discovery/news/2020/biomaterials-discovery...
 
Description Indo-UK Webinar on Secondary Ion Mass Spectrometry (ToF and 3D OrbiSIMS) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact This seminar was undertaken by David Scurr and coordinated by The University of Nottingham and Panjab University, Chandigarh, India. It covered an introduction to ToF and 3D
OrbiSIMS theory and instruments (capabilities and applications). A question and answer session was held afterwards focusing upon the applications of SIMS.
Year(s) Of Engagement Activity 2020
 
Description Invited talk at SIMS 23 (International SIMS meeting) - Minneapolis, USA, September 18-22, 2022 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Research dissemination regarding areas of progress made with the 3D OrbiSIMS instrument, highlighting some key findings.
Year(s) Of Engagement Activity 2022
 
Description Keynote presentation invite to 41st BMSS conference - Sheffield (UK) - 8th - 9th Sept. 2021 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other audiences
Results and Impact Keynote presentation at the 41st British Mass Spectrometry Society on the 08/09/21 attended by >100 participants. .
Year(s) Of Engagement Activity 2021
URL https://www.bmss.org.uk/41st-bmss-annual-meeting/
 
Description Morgan Alexander was an invited speaker at the 10th French-speaking TOF-SIMS users meeting 2021 (Online) 23 - 25 MARCH 2021 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Morgan Alexander was an invited speaker at the 10th French-speaking TOF-SIMS users meeting 2021 (Online)
23 - 25 MARCH 2021
Title: Introduction to the 3D Orbi SIMS with application examples from an academic lab
Year(s) Of Engagement Activity 2021
 
Description Pint of 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 JT was on the publicity team for Pint of Science 2016-17 and is a co-organiser for Pint of Science 2017-18.
Year(s) Of Engagement Activity 2017
 
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 13,000 attendees to the week long summer science event held in London. The grant developed and showcased 3 activities and 2 animated videos to the general public and media, with articles being published in the Times and the Telegraph as well as a slot on BBC Health Check regarding our work.

Telegraph Article - "New 'bacteria-phobic' material could stop the spread of superbugs in hospitals" https://www.telegraph.co.uk/news/0/new-bacteria-phobic-material-could-stop-spread-superbugs-hospitals/

The Times article - "New catheter cuts infection by keeping bacteria 'happy'"https://www.thetimes.co.uk/article/new-catheter-cuts-infection-by-keeping-bacteria-happy-t5h7rwdbs

BBC Healthcheck https://www.bbc.co.uk/sounds/play/w3csy9jy

In 2020 the Royal Society Summer Science Exhibition was ran as a virtual event.
Year(s) Of Engagement Activity 2019,2020
URL https://www.nottingham.ac.uk/research/groups/biomaterials-discovery/royal-society-summer-science-exh...
 
Description Wonder 
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 We invited families to explore the huge range of research and teaching activities that celebrated the Year of the Periodic Table. Activities had a fun link to the elements and ranged from art workshops through to science experiments.
Year(s) Of Engagement Activity 2019
URL https://www.nottingham.ac.uk/wonder/