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.

Publications

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Dundas A (2021) A new particle mounting method for surface analysis in Surface and Interface Analysis

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Hammann S (2020) Mechanisms of lipid preservation in archaeological clay ceramics revealed by mass spectrometry imaging. in Proceedings of the National Academy of Sciences of the United States of America

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Ruiz-Cantu L (2021) Bespoke 3D-Printed Polydrug Implants Created via Microstructural Control of Oligomers. in ACS applied materials & interfaces

 
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.
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,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 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 09/2017 
End 09/2022
 
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 03/2019 
End 03/2024
 
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 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 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 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"
The Times article - "New catheter cuts infection by keeping bacteria 'happy'"

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/