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.
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.
Flow cytometry
The Cancer Sciences Flow Cytometry Facility is a joint facility between the School of Cancer Sciences and CRUK Scotland Institute, providing access to a wide range of cutting-edge Flow Cytometers to the school, whole University community and to external users.
Our instruments can be used for:
- multicolour immunophenotyping
- olation of rare cell populations
- identification and isolation of fluorescent proteins in transfected cells
- cell cycle analysis
- apoptosis
- proliferation assays and many more functional assays.
Our team of highly trained specialists can provide as much technical support as required:
- training and advice on experimental design
- panel design
- sample preparation
- controls
- optimisation and troubleshooting
- support in data analysis and interpretation.
Wolfson Wohl Cancer Research Centre
BD LSR Fortessa Conventional Analyser
The Fortessa has five lasers: UV (355nm), Violet (405nm), Blue (488nm), Yellow/Green (561nm) and Red (640nm) which allow up to 18 fluorescent parameters to be simultaneously measured as well as forward scatter and side scatter.
The inclusion of a 561nm laser allows detection of red fluorescent proteins and better excitation of PE fluorochromes, utilising this laser reduces spillover and compensation with other channels.
Attune NxT Conventional Analyser
The Attune NxT has four lasers: Violet (405nm), Blue (488nm), Yello/Green (561nm) and Red (638nm). It can measure 14 fluorescent parameters simultaneously as well as forward scatter and side scatter.
The Attune NxT includes an autosampler capable of processing 96- and 384-well plates. Utilising innovative acoustic technology, this instrument experiences less clogging and can achieve a sample rate of up to 1 ml per minute.
SONY ID700 Spectral Analyzer
The ID7000 is a spectral cell analyser which has five lasers: UV (355nm), Violet (405nm), Blue (488nm), Yellow/Green (561nm) and Red (637nm) and 147 detectors. This instrument is different to our other, conventional cytometers as it doesn’t use physical filter sets but rather collects all the emitted light and unmixes the spectral signatures of individual fluorochromes. Whilst this is a different to conventional cytometry, the software is intuitive for experiment setup, acquisition, spectral unmixing and autofluorescence extraction.
The analysis software is free to all users of the instrument but requires a PC and 16GB of RAM. This software is not compatible with Apple.
The ID7000 has an autosampler which allows walk away analysis once settings have been optimised. Samples can be in 96 well plates, 384 well plates or 5ml tubes (FACS tubes).
As we dive deeper into our cells in more detail, the number of parameters increases, and with proper advice and experimental design, panels of over 40 fluorochromes are now possible.
BD Aria III Cell Sorter
The BD FACSAria III is a high-speed cell sorter which has four lasers: Violet (405nm),Blue (488nm), Yellow/Green (561nm) and Red (640nm) and can detect 16 fluorescent parameters plus forward scatter and side scatter, allowing for high purity, multicolour sorts.
The Aria can sort a range of particle sizes into a maximum of four separate populations, into FACS tubes, Eppendorf’s, cell culture plates or 15ml tubes.
BDFACS Aria Fusion Sorter
The BD FACSAria Fusion is a cell sorter which is located within a Class II biosafety cabinet allowing for sorting of containment level 2 samples.
The Fusion has three lasers: Violet (405nm), Blue (488nm) and Red (640nm) and can detect 11 fluorescent parameters plus forward and side scatter.
The Fusion can sort a range of particle sizes into a maximum of four separate populations, into FACS tubes, Eppendorf’s, cell culture plates or 15ml tubes.
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.
Paul O'Gorman Leukaemia Research Centre
BD Aria III Cell Sorter
The BD FACSAria III at POG is a cell sorter which has four lasers: Violet (405nm), Blue (488nm), Yellow/Green (561nm) and Red (633nm) and can detect 15 fluorescent parameters plus forward scatter and side scatter.
The Aria can sort a range of particle sizes into a maximum of four separate populations, into FACS tubes, Eppendorf’s, cell culture plates or 15ml tubes.
Paul O'Gorman (POG) is a containment level 2 laboratory meaning that CL2 samples can be sorted on this Aria.
BD Canto II Analyser
The Canto has three lasers: Violet (405nm), Blue (488nm) and Red (633nm) and can detect 8 fluorescent parameters plus forward scatter and side scatter.
POG is a containment level 2 laboratory meaning that unfixed CL2 samples can be analysed on the Canto.
Other resources and information
Health and Safety
All users using the facility must have completed the Health & Safety moodle courses prior to commencing work. We also advise reading the School of Cancer Sciences Heath & Safety manual.
The purpose of risk assessment is to help you identify the significant risks of your work to ensure that control measures are in place to protect people against harm so that the work can be carried out safely.
The Health and Safety at Work Act and the Management of Health and Safety at Work Regulations require a suitable and sufficient assessment of the risks to the health and safety of workers.
You are legally required to assess the risks in your work so that you can put in place measures to control the risks. The law does not expect you to eliminate all risks, but you are required to protect people as far as reasonably practicable by controlling the risks of your work.
The facility has a general RA for the instruments and a COSHH RA for the reagents provided for the instruments, but users are responsible for the RA’s associated with their work. The flow team can request access to these at any point. We can help with your RA’s if required.
Acknowledgement of the Facility in Publications
Acknowledgement of the facility in publications and presentations can be used as an identifiable metric to evaluate the support provided.
In line with the SCS Fair Publication Policy, we request that users ensure that a relevant acknowledgement is included in the relevant section of their manuscript.
Facility Acknowledgement: As a minimum, all publications resulting from the use of the instruments should acknowledge the facility, e.g. ‘The authors gratefully acknowledge the Cancer Sciences Flow Cytometry Facility for their support and assistance in this work’
Individual Acknowledgement: Where users have also had significant help from a particular member of the flow team. This should be considered where a staff member has exceeded standard assistance and furthered the research through experiment design, optimisation, data analysis or provided specialist technical troubleshooting. this staff member should be acknowledged by name, alongside the facility if applicable, e.g. ‘the authors thank *** of the Cancer Sciences Flow Cytometry Facility for their support & assistance in this work’
Authorship: In cases where a staff member has made a substantial intellectual contribution, over and above routine techniques. (e.g. Development or adaptation of protocols to suit samples or materials, (re)designing experiments, suggested a successful alteration of the protocol, extensive data acquisition, analysis and interpretation).
If you are uncertain about co-authorship or have any questions or concerns about this, please discuss this issue with the team in advance.
Please send a preprint of the paper, or an e-mail including the reference information for the publication, to the facility (scs-cruk_si-flow_facs_team@lists.cent.gla.ac.uk) and where appropriate the individual team member.
We also politely request that you remember to acknowledge use of the Cancer Sciences Flow Cytometry Facility in your presentations and cite the facility in grants, as appropriate.
Preparing Flow Cytometry Data for Publication
The flow team are happy to review flow cytometry data prior to publication, contact them directly for assistance.
MIFlowCyt (Minimum Information about a Flow Cytometry Experiment) establishes criteria for recording and reporting information about the flow cytometry experiment overview, samples, instrumentation and data analysis.
There is more information around this in our teams channel, including how to prepare MIFlowCyt and information on our instruments.
Data Storage Policy
Users are responsible for their own data files.
We recommend that you export your data immediately after sample acquisition.
Once data is successfully exported and backed up, we ask that you remove it from instrument software and from computer storage (i.e. desktops).
The Flow team will routinely delete data from the instruments, there will be fair warning prior to this.
In the event that you or a colleague are unable to back up your data due to exceptional circumstances please contact the Flow team.
Data Analysis Software
FlowJo: Available for both PC and Mac. Capable of analysing the FCS data from all our instruments. The facility has some dongles which can be signed out, the MVLS SRF flow cytometry core has signed up for FlowJo portal which we can join. If you would like more information regarding pricing/joining contact the Flow team.
Omiq: Cloud based platform which bridges machine learning and analytical pipelines with classical analysis. Capable of analysing the FCS data from all our instruments.
FCS Express: Capable of analysing the FCS data from all our instruments.
Cytobank: Cloud based platform, capable of analysing the FCS data from all our instruments.
Crusty: A free web tool for clustering analysis.
This is not an exhaustive list, contact the Flow team for further information.
New Users and External Users
New Users (Dropdown)
Request training and to book instruments on PPMS.
Contact the flow team, scs-cruk_si-flow_facs_team@lists.cent.gla.ac.uk or Jennifer.Cassels@glasgow.ac.uk, Yi-Hsia.Liu@glasgow.ac.uk, t.gilbey@crukscotlandinstitute.ac.uk with any queries, training or induction requests.
Users based in the CRUK Scotland Institute will need to request access to the Wolfson Whol and will need a University card to do so. Contact the flow team for instruction on this.
External Users
The Cancer Sciences Flow Cytometry Facility is happy to welcome users from other areas within the University, from other institutions and from external companies.
We understand that these users may require extra assistance and support in their flow cytometry experiments.
Please contact the flow team to discuss your requirements , scs-cruk_si-flow_facs_team@lists.cent.gla.ac.uk or Jennifer.Cassels@glasgow.ac.uk