The Cellular Imaging Core Facility (CICF) is a major hub for microscopy in Oxford, providing imaging solutions from the organismal to the molecular level. The CICF is much more than a point of use facility. In addition to excellent quality tailored user training and ongoing support on all systems covering a wide range imaging modalities, we also offer advice and guidance throughout each stage of imaging projects. We strongly encourage users come to the core at the project conceptualisation stage, we then work with them through planning, sample preparation, imaging, data analysis, figure preparation and finally publication.
Evident SoRa Super-Resolution Spinning Disc with High-Content Imaging and Analysis
The Olympus SpinSR SoRa is a spinning disc confocal microscope equipped with software for both standard microscopy experiments and high content multiwell plate assays. The Yokogawa CSU-W1 SoRa spinning disc unit features both a 50um pinhole disc for confocal imaging, and a SoRa super-res disc, allowing you to switch imaging modalities at the touch of a button. This enables rapid acquisition of large areas of interest, even when including z stacking. The system is very sensitive due to the camera based detection. This in combination with full co2, humidity and temperature controlled environment allows rapid acquisition of dynamic events, whilst maintaining long term viability. The microscope is also equipped with DIC for high contrast brightfield acquisition.
Excitation - diode lasers
405nm
488nm
561nm
647nm
SpectrX for advanced widefield
Objectives
4x
10x 0.4NA
20x 0.8NA Oil
40x 0.95NA Air
40x 1.4NA Oil
60x 1.2NA Sillicone
60x 1.4NA Oil
Detection
2x Hammamatsu ORCA Fusion back thinned cameras
Filter sets for DAPI/Alexa488/Alexa568/Alexa647 including single and multipass options
Software
The Evident VS200 is a widefield slide scanner, designed for rapid imaging of a wide range of samples. Featuring an automated robot loader, the VS200 has the ability to scan up to 210 slides in a single run. The system can image transmitted light brightfield, polarised light, and fluorescence. For fluorescence, the slide scanner features 5 imaging channels (DAPI/GFP/RFP/CY5/NIR channels). The system is equipped with 2x, 4x, 10x, 20x and 40x air objectives, and a 60x auto immersion oil lens. The system has two full software licenses (one for acquisition, one for offline data analysis), and includes AI sample detection for fast workflows, minimising user input.
Access to a range of analysis software packages including Arivis, Imaris, Zen, scanR, CellProfiler, and FIJI installed on ultra-high spec processing PC's allows quantification of large, multidimensional datasets. Support is offered to generate analysis pipelines utilising the latest advances in the field such as AI (pre trained networks such as cellpose, and tools to generate bespoke machine and deep learning networks), automated neurone tracing and live cell tracking. This, In combination with the imaging systems available in the core, allows truly high throughput imaging and analysis workflows to be created for users.
The SP8 Fast Lifetime Contrast (FALCON) is a point scanning confocal microscope with lifetime imaging and analysis built in to the acquisition software. The system has a fully automated stage and 7 fixed laser lines, allowing fast acquisition of multichannel, multi-position experiments. The microscope also features a pulsed 440nm laser for lifetime experiments. This in combination with a high sensitivity single photon time-gated counting spectral detection array and full temperature, humidity and co2 incubation allows a diverse range of experiments to be performed on the microscope.
Excitation
405nm, 458nm, 488nm, 495nm, 514nm, 561nm, 633nm
440nm pulsed for FLIM
Objectives
10x 0.3NA
20X 0.7NA
60x 1.3 water immersion
100X 1.4NA oil immersion
Detection
4x Hyd
1x pmt
Software
LASX with FALCON module
The Leica SP8 is a point scanning confocal microscope which features a white light laser for excitation. This allows selection of any excitation wavelength within the range of 470-670nm, which in combination with the standard fixed lines on the system allows careful excitation of almost all commonly used fluorophores. The SP8 also features a unique spectral detection array, allowing careful separation of emission from multiplex assays.
Excitation
405nm, 458nm, 488nm, 495nm, 514nm, 561nm,
470-670nm White light laser
Objectives
10x 0.3NA
20X 0.45NA
40x 1.3NA oil
60x 1.4NA oil
Detection
2x PMT
2x HyD
Spectral detector array allows flexible selection of emission band
Software
LASX for acquisition and basic analysis and 3D Rendering
Picoquant for FLIM experiments
Fully automated upright and inverted Leica microscopes each equipped with a monochrome camera for high sensitivity fluorescence assays, and a colour camera for histopathological stains. Both systems feature automated stages which in combination with the navigation and multi-well plate modules in the software and fast filter wheels allows rapid acquisition of large fields of your sample in multiple channels.
Excitation
Individual Filter cubes for DAPI/GFP/txRED/CY5
Dual pass filter cube: CFP/YFP
Quad pass fast filter wheel; DAPI/GFP/RFP
Objectives
5X 0.12 NA
20X 0.4NA Long working distance
20x 0.8NA
40X 1.3NA Water immersion (not wavelength corrected in Z)
100X 1.4NA
Software
LASX
The Zeiss LSM900 is a point scanning confocal microscope, which in combination with the second generation Airyscan detector allows user friendly super resolution imaging without the need for complex sample preparation protocols. This system includes a fully motorised stage with piezo z drive for rapid acquisition of multiple positions, with the option to z stack and tile positions seamlessly. This system is also equipped with full incubation for live cell experiments including temperature, humidity and co2 control. In combination with the incredibly sensitive Airyscan detector, this allows long timelapse super-res imaging of cellular dynamics with minimal impact on cell viability.
Excitation - diode lasers
405nm
488nm
561nm
647nm
Objectives
2.5x
10x 0.35NA
20x 0.8NA
40x 1.2NA Water
63x 1.2NA Water
63x 1.4NA Oil
Detection
3x PMT
1x TPMT for transmitted light acquisition
1x Airyscan 2 Detector
Combination of variable sliding dichroics and fixed filters allow careful selection of emission range.
The Elyra PS1 is a widefield fluorescent microscope which combines TIRF and SMLM modalities in to a single system. Single molecule localisation microscopy uses the phenomenon of fluorophore ‘blinking’ to push the resolution limit of light microscopy down to the 30-40nm scale. TIRF imaging allows high resolution imaging of structures close to the coverslip with fantastic signal to noise ratio, and so is ideal for imaging structural proteins and targets which sit on the cell surface. This system features a dual camera setup, allowing rapid simultaneous multichannel acquisition.
Excitation
Diode lasers - 405nm, 488nm, 561nm, 647nm
Metal-halide lamp for widefield
Objectives
100X 1.45NA
60X 1.4NA
10X 1.35NA
Detection
2 X iXon Ultra EMCCD - back-illuminated 512 x 512
Software
ZEN Black
The Zeiss Microbeam Laser Capture Microdissection offers unique ‘laser catapulting’ technology. This involves using a high power UV laser to cut around regions of interest on tissue sections, down to the single cell level. The system then uses a defocused pulse of laser light to catapult the cut area in to the lid of a tube, which can then be taken for downstream analysis. The microscope comes equipped with a robomover automated sample capture system, which allows cuttings to be automatically directed to up to 96 different caps within a single experiment. Samples can be imaged and cut out using both colorimetric and fluorescent protocols.
Fluorescence
Objectives
Detection
