Expertise
Imaging Platforms
Keywords - Light Microscopy: Confocal microscopy | Superresolution microscopy | Widefield microscopy | TIRF | Live cell imaging | PALM | STORM | FCS/FCCS | Structured Illumination Microsocpy
Keywords - Electron Microscopy: Transmission EM | Scanning Transmission EM | Electron tomography | Scanning EM (FEG) | Cryo-SEM
The JIC Bioimaging facility has a super-resolution system (Zeiss Elyra PS1), a spinning disk confocal (Andor Revolution XD) and five point-scanning confocals (Zeiss LSM 780, 880 with Airyscan, Leica Stellaris and SP8) plus a good range of other wide-field and stereo light microscopes, all with digital cameras. A range of inverted and upright systems will cover most requirements. Image analysis workstations are also available.
The electron microscopes are cryo-capable and have field-emission guns. The TEM is a Thermo Fisher Talos F200C fitted with a Gatan OneView camera and will have a Falcon 4i detector in early 2023. There are two SEMs (Thermo Fisher Nova NanoSEM 450 and Zeiss Gemini 300), both with cryo-systems (Gatan and Quorum respectively). ETD, TLD (In lens), BSE, STEM and EDS detectors are available and we have Gatan 3View system with an OnPoint detector on the Zeiss SEM. There is a variety of specialist sample preparation equipment for biological EM such as a grid plunging device, various coaters, a high-pressure freezer, embedding machines and ultramicrotomes.
Applications
Keywords – Biological: Cell Biology | Developmental Biology | Plant Biology | Microbiology | Bio-materials | Food
Keywords - Physical Sciences: Bioengineering and biomaterials | Nanomaterials (nanoparticles, 2D nanomaterials, nanowires and nanotubes) | Polymers and organic electronics
Whilst our expertise is particularly strong in the two areas of high-resolution cryo-SEM and live-cell imaging using GFP technology, we support a wide range of specialist imaging techniques and can accommodate various types of samples, from whole tissues to nano-structure imaging.
All our confocal systems are equipped with multiple lasers (to excite most available dyes) and highly sensitive detectors. This enables detailed analysis of living cells. Using our in-house designed perfusion chambers, long-duration time-lapse observations can be performed over several days, to study developmentally regulated dynamic events.
Other advanced imaging techniques are available including electron tomography, single particle analysis, cryo-TEM, super-resolution (STORM, PALM and SIM), FLIM and FCS. We also routinely prepare resin-embedded material, perform immuno-gold labelling and image negative-stained viruses and nano-particles in the TEM.
Sample Preparation
Keywords - Biological: Resin embedding | Ultrathin sectioning | Serial sectioning | Immunolabelling | Plunge Freezing | High Pressure Freezing | Freeze substitution | Critical Point Drying | Freeze fracture | Metal coating | Negative stain | Photoconversion | PLT | Immunofluorescence | Cryo-TEM | Cryo-SEM | Cryostat sectioning | Vibrating microtome | Enzyme histochemistry | Live Cell
Samples can be prepared by a variety of techniques for TEM such as cryo-grid plunge using the Vitrobot, high-pressure freezing and freeze-substitution or by chemical fixation and resin embedding, including the PLT method. Negative staining and other techniques such as PATAg staining and immuno-gold labelling are also available. The 200kV TEM has a FEG electron source and can perform electron tomography with high and dual tilt holders. For SEM, cryo-techniques are a speciality at JIC but critical point drying is also routinely used. We offer plunge freezing as a routine cryo-SEM method, but high-pressure freezing or impact freezing for high resolution freeze-fracture work is also available. We have sputter coaters, carbon coaters and a glow discharge machine. In addition to ultrathin resin sectioning for EM, we can also cut semi-thin resin sections or wax, vibratome or cryostat sections for light microscopy.
Data Analysis
Keywords - Software: ImageJ | FIJI | Amira | Matlab | AutoQuant | Metamorph | AxioVision
Image data can be analysed in-house using dedicated workstations. Commercial software from Leica, Zeiss, Molecular Devices and Media Cybernetics are available offline. There is a dedicated workstation for super resolution (SIM / PALM / STORM) image processing. For 3D visualisation and analysis, we use either FEI Amira or open source VolViewer.
Shared Access
The Bioimaging facility at the John Innes Centre (JIC) is one of a number of research facilities within JIC and across the partner institutions that comprise the Norwich Research Park (NRP). Many of these facilities (including Bioimaging) are available for use by NRP institutions as well as the wider community external to the NRP.
In common with all the NRP shared facilities, the Bioimaging facility at JIC uses an FEC model to help recover costs. An on-line booking system is available for internal use but external clients should contact the facility manager to discuss their needs. Comprehensive training is provided for those people wishing to use specific Bioimaging equipment without supervision.
Although JIC’s primary research focus and reputation concerns plant and microbial sciences, the Bioimaging team has been involved with an increasing number of applications outside the scope of such research. The team welcomes enquiries from anyone interested in using the imaging facility.
Funding
The majority of the JIC’s funding is won in open competition from funding agencies worldwide, with more than 50% coming from UK government sources. BBSRC provides a large proportion of the JIC’s funding in the form of responsive mode funding and strategic funding for the Institute Strategic Programmes.
The JIC also holds grants from the European Research Council and charitable sources.