Imaging dynamic biological systems across scales

Bioimaging has become one of the leading approaches to study biological processes in cells, tissues and whole organisms with the help of light and other non-invasive methods.

In particular, recent breakthroughs in fluorescence microscopy have revolutionised the way biomedical research is conducted, allowing the direct observation of biological processes in living cells, tissues and organisms with unprecedented spatial and temporal resolution.

The expertise and equipment available at the COMPARE Advanced Imaging Facility span across multiple scales and imaging modalities, from the observation of individual proteins diffusing and interacting on the surface of living cells to the visualisation of complex cell organisation in large tissue samples.

We combine these with innovative probes such as small fluorescent molecules and FRET/BRET sensors developed by our word-leading PIs, in-house hardware design, and  development of novel computational approaches based on AI and machine learning.

Our researchers and support staff have extensive experience in applying these methods to a wide range of biomedically relevant questions, such as clarifying the mechanisms of cell-cell communication and receptor signalling, investigating the pathogenesis of metabolic, inflammatory and cardiovascular diseases, or developing innovative pharmacological therapies and diagnostic markers.

 

Image 3 230x230

Single Molecule Localisation Microscopy (dSTORM) showing 3D structure of the microtubule network in CHO cells.

(Image courtesy of Dirk-Peter Herten)

Image 4 234x231

SIM image of platelets spreading on collagen fibres, labelled for F-actin (cyan) and GPIbα receptor (magenta).

(Image courtesy of Natalie Poulter)

Image 5 230x230

Volumetric imaging of optically cleared whole mouse embryo (E15.5) seen in autofluorescence on the Ultramicroscope II.

(Image courtesy of Werther Vecchiato)

 

Image 1

Maximum intensity projections of actin (green) and tubulin (red) in an extension of a primary megakaryocyte cell. Cytoskeletal dynamics  during proplatelet formation was monitored every minute in 3D on the Lattice Light Sheet.

(Image courtesy of Malou Zuidscherwoude)

Image 2 245x174

Single-molecule receptor-arrestin interactions: tracking individual molecules as they diffuse and interact in living cells with high spatiotemporal resolution (SMLM), providing information on molecular diffusion, interactions and nanoscale arrangement.

(Image courtesy of Davide Calebiro)

 

Our Technologies and Applications

List of microscopy techniques with their associated use and systems
 TechniqueUse  Systems
Super-resolution microscopy 
Structured Illumination Microscopy (SIM) Visualisation of sub-cellular structures (live and fixed cell) at twice the resolution of conventional microscopy (~ 160 nm) Nikon N-SIM
Single-molecule Localisation Microscopy (SMLM) Provides quantitative information on complex populations of molecules, including their nanoscale arrangement, diffusion and interactions in live cells with unprecedented resolution (~20 nm and 20 ms) Custom 4-camera TIRF (Cairn)
Stochastic Optical Reconstruction Microscopy (STORM) Super-resolution imaging of molecules within cells and tissues, labelled with antibodies or photoswitchable probes. Resolution ~ 10-20 nm Nikon N-STORM

Nanoimager (ONI)
Light sheet microscopy  
Lattice light sheet (LLS) Non-destructive imaging of living samples in three dimensions over time with unparalleled spatiotemporal resolution (~160 nm) 3i Lattice Light Sheet
Selective Plane Illumination Microscopy (SPIM) Provides precise information on cellular/organelle dynamics in live cells, cleared tissues and small organisms, with 3D isotropic resolution 3i Marianas Light Sheet 
Large Volumetric Imaging Light Sheet 3D imaging of entire biological systems in large optically cleared samples (e.g. whole brain,  organoids, tissue slices) with cellular resolution Ultramicroscope II (Miltenyi)
Oblique Plane Light Sheet – coming soon! Fully automated, remotely accessible high content light sheet microscopy system – we’re now looking for pilot projects, please ask for further information Custom 5-camera OPLSM
Widefield microscopy    
Calcium imaging /cellular Kinetics  Receptor activation or the production of second messengers such as calcium or cyclic AMP in living cells  Ultrafast FRET/calcium imaging microscope (Cairn)
 FRET Protein-protein interactions in living cells Ultrafast FRET/calcium imaging microscope (Cairn)

Custom 4-camera TIRF (Cairn)
Multichannel Fluorescence microscopy Automated imaging applications such as time-lapse imaging, multiwell plate scanning, image stitching and tiling, and cell counting Ultrafast FRET/calcium imaging microscope (Cairn)

Thermofisher EVOS FL