Euro-Bioimaging

UNITED KINGDOM

The UK Node

The UK Node offers open-access to a wide range of advanced biological imaging techniques including correlative, multi-modal, high-content and super-resolution. It is a multi-sited national infrastructure hosted by the following seven leading institutions spread across the UK: Edinburgh Super-Resolution Imaging Consortium (ESRIC), the Francis Crick Institute, King’s College London, Liverpool University, Octopus at Harwell, Oxford Brookes University and York University. They all offer state-of-the-art imaging equipment, expertise, training and image data services. The technologies can be applied to a wide range of fundamental and translational research projects at molecular to cellular resolutions, in single cells to 3D, in vitro models and whole organisms.

The Euro-BioImaging UK Node Manager acts as the single point of contact who can direct potential users to the relevant imaging technology and site to undertake their experiments. We welcome applications from all biological and biomedical research disciplines. Projects will be prioritised based on technical feasibility and scientific merits. Support will be provided for the entire experimental pipeline including (where appropriate) initial user consultation, experimental planning, full hands-on training, assistance with sample preparation, gathering highest quality imaging data and data analysis.

Specialities and Expertise of the Node

ESRIC: SIM, dSTORM, PALM, spt-PALM, SRRF, STED, FLIM-FRET, TIRF, Spinning Disk Confocal

ESRIC comprises open access advanced imaging facilities specialising in super-resolution techniques. As well as cutting-edge equipment, we bring together a broad spectrum of knowledge and expertise in extending the boundaries of biological imaging beyond the diffraction limit. Current technologies include dSTORM/PALM/DNA-PAINT on Nikon N-STORM and Olympus Cell Excellence systems, gSTED, Tau STED and FLIM-FRET on a Leica SP8 with FALCON FLIM, Structured Illumination Microscopy on a Nikon N-SIM /SoRa hybrid system, and SRRF on an Andor Dragonfly spinning disk. Our equipment enables us to image objects in 3D at super-resolution in up to 4 colours, with resolutions ranging from 120nm down to 10nm.

Francis Crick Institute: Lightsheet microscopy (from live organoids and embryos to fixed whole mouse organs), High Resolution Episcopic Microscopy (HREM), Optical Projection Tomography (OPT)

We offer two methods for imaging thick samples, Lightsheet microscopy and High Resolution Episcopic Microscopy (HREM), with support for the entire experimental workflow, including sample prep, clearing, image analysis, and data management.

Our Lightsheet systems cover two distinct application spaces: Larger fixed samples (10 - 20 mm) can be imaged with the LaVision Ultramicroscope whereas smaller living or fixed samples (50 - 500 µm) can be imaged with either the Luxendo MuVi or Viventis LS1. The Ultramicroscope can be used in combination with antibody labelling and tissue clearing to image many sample types, including whole mouse organs, later-stage embryos, tumors, and tissue sections. In contrast the MuVi and LS1 are suitable for live imaging of small samples such as spheroids, organoids, and early-stage embryos. The LS1 offers higher throughput whereas the MuVi can image thicker samples, and has been adapted with a pulsed IR laser for photo-stimulation.

HREM is a simple block-face approach for imaging tissue and organ morphology that does not require endogenous tissue fluorescence or any form of antibody labelling. It was originally developed to characterise embryonic lethal mouse mutations and has been used to study cardiovascular, skeletal, and neurological defects in a wide variety of mouse models (eg: Vanyai et al, Development 2020.)

King’s College London: STORM ,SIM, Lightsheet, FLIM, CLEM,Volume EM, LA-ICP-MS, Raman

At the atomic and nano scales, we support volume EM and CLEM workflows, cryo SEM and cryo TEM and are experts in freeze fracture. We provide cryo-focused ion beam SEM and are developing a complete pipeline from live cell to cryo ET leading to sub-tomographic averaging of proteins in situ.

Moving down the resolution scale, we provide access and support in, super-resolution and low-toxicity live cell imaging (N-STORM 5.0 and SoRa, plus single and multi-photon lightsheet capability), as well as high-content and throughput spinning disc (Opera Phenix).

Extending beyond optical imaging, we deliver analytical multi-modal imaging in the form of 2D or 3D spatial quantitative elemental and molecular imaging of both endogenous elements and metal tagged antibodies in cells and tissues using a multiplexing methodology whilst identifying associated lipids and biomolecules in a label free approach. Routine LA-ICP-MS (Laser Ablation Inductively Coupled Plasma Mass Spectrometry) has been integrated with DESI (Desorption electrospray ionisation)-MS and Raman Spectroscopy as a single unique correlative workflow. Together with integration of IR, this workflow will enable high resolution spatial mapping of lipid, nucleic acids and protein phosphorylation.

Liverpool: Automated Live Cell Imaging, STORM, PALM, Bio AFM-TIRF, SIM2, Lightsheet, Cytometry, volume EM, TEM, MicroCT, Photoacoustic

We provide an integrated ensemble of imaging modalities in one place for imaging across scales from single molecule to cells and model organisms. Example projects include cell biology, surface science, biochemistry, and microbiology. Of particular note is our unique Bio AFM-TIRF imaging system for atomic force microscopy combined with dual fluorescence total internal reflection. We support the entire experimental workflow for each technique with dedicated staff, and we use an OMERO server for data management.

Octopus: MINFLUX, CLEM with superresolution and cryoFIB-SEM, SIM, STORM/PALM, STED, Lightsheet, Multiphoton, FIB-SEM, FLIM/PLIM, Optical Trapping, TIRF, Raman, Single Molecule Tracking

We offer multiple microscopy techniques on a suite of commercial and custom-built systems, including MINFLUX, light sheet, single molecule localisation microscopy (PALM/STORM, including cryo-STORM), STED, single molecule methods including tracking and Fluorophore Localisation Imaging with Photobleaching (5 nm precision), confocal (including two photon and FRET/FLIM), optical trapping, and cryo focused ion beam SEM.

The microscopes are co-located with extensive preparation facilities including laboratories for biochemistry, cell biology, and chemistry. Facility users have access to a dedicated support team including microscopists, cell and molecular biologists, chemists, and data analysis experts. Users are provided with assistance at all stages of the experiment, including sample preparation and labelling, collection of data, and data analysis and interpretation. Custom data analysis packages are available and can be tailored to the requirements of a particular experiment. Accommodation and catering facilities are available on campus.

Oxford Brookes: SBF-SEM, SEM (tomography), CLEM, Cellular Electron Tomography, High Pressure Freezing

We focus on providing volumeEM and CLEM workflows, including dual axis serial section cellular electron tomography and serial block face-scanning electron microscopy (SBF-SEM). We can work with fixed or high pressure frozen samples and have extensive expertise in eukaryotic parasite work and insect vectors. The centre is equipped with CATII culturing facilities within the Centre, as well as a SAPO licence to support these projects.

York: SIM, Multiphoton, Spatial Transcriptomics (GeoMX), Phase Imaging /Holography, Confocal, Slide Scanner, cryoTEM, cryoSEM

Current technologies include PALM/STORM/SIM2 via the Zeiss ELYRA 7, Spatial-omics via Nanostring GeoMX-DSP and 10X Visium, Label-free imaging via PhaseFocus LiveCyte and Tomocube and multiphoton imaging via Zeiss LSM980 AiryScan2 along with the more routine confocal, slide scanning and scanning and electron microscopy. The team have a wide breadth of biological expertise spanning plant sciences through to biomedicine, and are familiar with most sample types and sample preparation methods.

Additional services offered by the Node

Project planning and management
Wet lab
Cell culture facilities
Methodological setup
Facility induction
Technical assistance to prepare experiments and run instruments
Workstations
Data storage
Image acquisition
Image processing and analysis

Probes

All sites within the EuBI UK Node operate within leading edge academic institutions and as such, many different types of probes, including plasmids, antibodies and chemical reagents are available within individual researcher labs. Additionally, a number of facilities maintain their own in-house reagents, including pre-labelled antibodies for super-resolution (STORM) and spatial transcriptomics.

Other model organisms

All sites within the EuBI UK Node operate within leading edge academic institutions that host labs working with a broad range of different model organisms, including zebrafish, Drosophila and c.elegans. Facilities to support users working with such organisms would therefore be possible, subject to discussions and agreements with the appropriate academics.

High biological safety level

BSL2 facilities are available within a number of the laboratories at the host institutions for the EuBI UK Node facilities and can be made available to users following consultation with the site leads. Of note, Oxford Brookes holds a SAPO license for culturing Trypanosoma brucei within the imaging facility itself.

Instrument Highlights

ESRIC: dSTORM/PALM/DNA-PAINT on Nikon N-STORM and Olympus Cell Excellence systems, gSTED, Tau STED and FLIM-FRET on a Leica SP8 with FALCON FLIM, SIM on a Nikon N-SIM /SoRa hybrid system, and SRRF on an Andor Dragonfly spinning disk. Our equipment enables us to image objects in 3D at super-resolution in up to 4 colours, with resolutions ranging from 120nm down to 10nm.

Francis Crick Institute: Our Lightsheet systems cover two distinct application spaces: Larger fixed samples (10 - 20 mm) can be imaged with the LaVision Ultramicroscope whereas smaller living or fixed samples (50 - 500 µm) can be imaged with either the Luxendo MuVi or Viventis LS1. The Ultramicroscope can be used in combination with antibody labelling and tissue clearing to image many sample types, including whole mouse organs, later-stage embryos, tumors, and tissue sections. In contrast the MuVi and LS1 are suitable for live imaging of small samples such as spheroids, organoids, and early-stage embryos. The LS1 offers higher throughput whereas the MuVi can image thicker samples, and has been adapted with a pulsed IR laser for photo-stimulation.

King’s College London: At the atomic and nano scales, we support volume EM and CLEM workflows, cryo SEM and cryo TEM and are experts in freeze fracture. We provide cryo-focused ion beam SEM and are developing a complete pipeline from live cell to cryo ET leading to sub-tomographic averaging of proteins in situ.

Liverpool: We provide an integrated ensemble of imaging modalities in one place for imaging across scales from single molecule to cells and model organisms. Example projects include cell biology, surface science, biochemistry, and microbiology. Of particular note is our unique Bio AFM-TIRF imaging system for atomic force microscopy combined with dual fluorescence total internal reflection. We support the entire experimental workflow for each technique with dedicated staff, and we use an OMERO server for data management.

Octopus: We offer multiple microscopy techniques on a suite of commercial and custom-built systems, including MINFLUX, light sheet, single molecule localisation microscopy (PALM/STORM, including cryo-STORM), STED, single molecule methods including tracking and Fluorophore Localisation Imaging with Photobleaching (5 nm precision), confocal (including two photon and FRET/FLIM), optical trapping, and cryo focused ion beam SEM.

Oxford Brookes: We focus on providing volumeEM and CLEM workflows, including dual axis serial section cellular electron tomography and serial block face-scanning electron microscopy (SBF-SEM). We can work with fixed or high pressure frozen samples and have extensive expertise in eukaryotic parasite work and insect vectors. The centre is equipped with CATII culturing facilities within the Centre, as well as a SAPO licence to support these projects.

York:

Left: Cyanobacteria imaged on the Zeiss Elyra7, Luke Mackinder Group. Right: Algae imaged on the Tomocube, Luke Mackinder Group

Contact details