Cellular Analysis
We offer support in:
- Light microscopy (basic imaging, live-cell imaging, confocal microscopy)
- Flow Cytometry
- Image analysis support
We work very closely with the CRUK Scotland Institute ‘Advanced Imaging’ Facility (BAIR), who provide in-depth support for several aspects of the microscopy and FACS facilities.
Light microscopy
School of Cancer Sciences houses a range of microscopes from basic analysis in tissue culture suites through to more sophisticated fluorescence confocal microscopes.
Agilent Seahorse
The Agilent XFe96 offers real-time cellular metabolic analysis. Whilst each cell contains over 20,000 genes, 200,000 proteins and thousands of pathways, you can't measure everything in a cell at once, but you can measure the energy that drives them.
Agilent offers Seahorse XF cell analysis technology solutions that detect discrete changes in cellular bioenergetics in real-time. Real-time cell metabolic measurements provide a clear window into the critical functions driving cell signalling, proliferation, activation, toxicity and biosynthesis.
Agilent Seahorse XF Analysers perform automatic, real-time measurements of oxygen consumption rate (OCR), proton efflux rate (PER) or extracellular acidification rate (ECAR), as well as ATP production rates of live cells in a multi-well plate, interrogating key cellular functions like mitochondrial respiration and glycolysis. The analysers also perform compound addition and mixing along with label-free analytical detection, providing a systems-level view of cellular metabolic function in cultured cells and ex-vivo samples.
ZEISS Axio Vert A1
Basic Axio Vert microscopes are supplied in all the tissue culture rooms and enable convenient acquisition of lower-quality images.
ZEISS Axio Observer A2
For better quality images, we have the Axio Observer, which can be used for widefield fluorescence microscopy, or for transmitted light microscopy and has been adapted for live-cell imaging
ZEISS LSM 780 Confocal Microscope
Specialised for capturing high resolution fluorescence images.
Airy-Beam Light Sheet Microscope (MSquared)
Light sheet fluorescence microscopy is an increasingly popular imaging technique for producing high contrast 3D volumetric images of intact biological specimens. This microscope can image at sub-cellular resolution over entire organisms, from >10mm to less then 1µm.
Olympus LV200 Bioluminescence Imaging System for imaging subcellular NanoBRET interactions
Highly specialised microscope system designed to measure drug:target interactions within a cell using “NanoBRET” technology (Bioluminescence Resonance Energy Transfer).
Enabling the detailed study of photosensitive cells and luminescence probes at high magnification, the LV200 bioluminescence imaging system’s built-in temperature control, humidity, and gas flow protect cultured cells and tissue slices during long-duration observation.
Optimised for both slides and cultured cells, as well as standard fluorescence microscope uses:
- LV200 Frame with motorised XY and focus
- pE300 LED light source for brightfield and fluorescence
- Transmission fluorescence filter sets for DAPI, GFP, RFP, Cy5 and like dyes
- Hamamatsu EMX2 High Sensitivity EMCCD camera
- Stagetop incubator with temperature and gas mixing control
- PC workstation with CellSens imaging software to control system
- 10x, 20x, 40x and 60x Oil immersion lenses
Opera Phenix / Screening Service
Opera Phenix Plus is designed for high-throughput high-content imaging assays, phenotypic screening, assays using complex disease models, such as live cells, primary cells and microtissues, and fast-response assays, such as Ca2+ flux.
IncuCyte
A live-cell imaging and analysis system that can be used to study cell behaviour over time. Used to perform a variety of experiments, including cell health and proliferation studies, chemotaxis and stem cell differentiation. This can be used in conjunction with a hypoxia cabinet, if required.
Flow cytometry
Our machines can be used for:
- Multicolour immunophenotyping
- Cell cycle analysis
- Identification and isolation of fluorescent protein-transfected cells
- Isolation of rare cell populations
- Apoptosis assays
- Functional assays
What we offer:
- Cell sorting
- FACS analysis
- Spectral FAC analysis
- FACS-RTT (Fluorescence-Activated Cell Sorting Radioligand Treated Tissue)
BDFACS Aria Fusion Sorter
The BD FACSAria™ Fusion Flow Cytometer combines sorting capabilities with best-in-class biosafety expertise for a comprehensive advanced cell sorter and biosafety solution. 14 colour high speed cell sorter with 3 lasers (Violet 405nm, Blue 488nm and Red 640nm)
The fully integrated biosafety cabinet meets emerging operator and sample protection requirements as well as many country standards for bioprotection. The instrument uses BD FACSDiva™ Software to efficiently control the setup, acquisition and analysis of flow cytometry data from the operator workstation.
FACS BDFACS Aria Sorter (POG)
The BD FACSAria cell sorter has a revolutionary design that has created a stable instrument with which to perform cell sorting.
This has been achieved by introducing a number of novel features: fixed-alignment cuvette flow cell; self-contained fluidics and; improved computer software. These advances have revolutionised, while at the same time simplified, the way cell sorting can be achieved.
Our cell sorter offers a number of powerful features:
- Three lasers - 407-nm Violet, 488-nm Argon, and 633-nm HeNe
- Multi-parameter analysis which allows up to 12 independent signals to be analysed at one time for a more detailed phenotypic analysis of cell populations
- Digital compensation to increase sensitivity of analysis
- Acquisition of up to 90,000 events/second
- Sorting of up to 25,000 events/second, with > 98% purity
- Collection of four cell samples concurrently to reduce the amount of sample required
- Sorting into a number of different collection devices, such as: microtubes, 5 ml and 15 ml tubes and 6, 12, 24, 48, 96, 384 well plates
- Isolation of single cells in tissue culture plates for cloning purposes
- Sorting of defined ratios of multiple cell types in a single-pass for experimental set-up
- Collection of rare cell events, due to the increase in sensitivity
- Temperature control of samples - the instrument offers temperature regulation (heating or cooling) of unsorted and sorted samples
- Aerosol management system for operator protection from potentially biohazardous aerosols
BD FACS CANTO II (POG)
Our FACS analyser, the BD FACSCanto II, has a similar fluorescent footprint to our FACSAria, but is tailored for analysis of specific biological properties of mixed cell populations from multiple samples.
SONY ID700 Spectral Analyzer
Sony ID7000 is a 5-laser instrument (355nm, 405nm, 488nm, 561nm, 660nm) with 147 detectors.
It is different to our conventional cytometers as it doesn’t use physical filter sets (so we don’t have a traditional configuration document) but rather it takes all of the light emitted and unmixes the spectral signatures of individual fluorochromes.
Whilst this is a different way of working the instrument has intuitive software for experiment setup, spectral unmixing and autofluorescence extraction.
The analysis software is free to all users of the system but runs on a PC and requires 16GB of RAM.
Acquired data must be unmixed at the instrument but thereafter can be analysed offline.
As we look in more detail at our cells, the number of parameters needed increases, and with the proper advice and experimental design, panels of over 40 colours are now possible.
The instrument has an autosampler and walk away analysis is truly possible once settings have been optimised.
Enquire about SONY ID700 Spectral Analyzer
Cytek Northen Lights Flow Cytometer
In addition to the FACS Hub, there is a Cytek Northen Lights Flow Cytometer located in the Translational Pharmacology & Immunology Laboratory, operating under GCP guidelines.
The Cytek Northern Lights™ flow cytometer, with its easy-to-use workflow, is a flexible and affordable full spectrum flow cytometry solution. The Northern Lights instrument incorporates Cytek’s powerful Full Spectrum Profiling™ (FSP™) technology in a one to three laser system, allowing for a wide array of new fluorochrome combinations without reconfiguring your system for each application.
Whether you are doing basic assays such as cell proliferation, apoptosis analysis, or other cell health applications, or multicolour immunophenotyping panels, the Northern Lights system meets all of your flow cytometry needs.
Hypoxia Incubator
There is an H35 Whitley Hypoxia Incubator in the laboratory on Level 1 in WWCRC. The hypoxia chamber allows precise control of oxygen, carbon dioxide, humidity and temperature, providing physiologically relevant conditions for cell culture, simulating low-oxygen conditions and the effect on biological systems. With the H35, oxygen levels can be controlled in 0.1% increments up to 20%; carbon dioxide in 0.1% increments up to 15%; and relative humidity up to 80%. This provides a great deal of flexibility for a range of research requirements.