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The Flow Cytometry Translational Technology Platform provides equipment and expertise in flow analysis, cell sorting and mass cytometry to cancer researchers at UCL.
Flow cytometry is a powerful tool that uses lasers to rapidly quantify the fluorescent and light scattering properties of single cells or particles in suspension. Cell components can be labelled with fluorescent dyes to allow different populations to be delineated from the bulk population. Dyes can be used to label and quantify DNA, epitope expression and also for functional assays including oxidative metabolism, apoptosis/viability, calcium flux and proliferation.
The Flow Cytometry Translational Technology Platform at the UCL Cancer Institute is equipped with the latest cutting-edge instrumentation with a BD Symphony A5 capable of measuring up to 28 fluorescent parameters including access to the latest fluorescent dye technology and a CyTOF XT Mass cytometer capable of measuring >50 parameters simultaneously per cell. The facility also has three 4-laser Fortessa analysers and three cell sorters, one of which is housed in a Class II Biosafety cabinet. Services provided by the facility include Cell Sorting, Mass cytometry and analyser training from beginner to advanced levels including high-dimensional data analysis.
The TTP is supported by two specialists and the facility manager with many years of cumulative experience. We provide expertise in experimental design for both sorting and analysis as well as technical assistance.
Booking
Following appropriate training, you will be given access to the relevant booking calendars on the Clustermarket Bookkit system.
Equipment
Analysers
- BD Fortessa X20
We have two Fortessa X20 analysers in the TTP. Both instruments are equipped with 4 lasers (Violet, Blue, Yellow-Green and Red) capable of measuring 16 parameters, providing a robust and flexible system for researchers in the CI. Full instrument training is provided to researchers by members of the TTP.
- BD LSR Fortessa C
The LSR Fortessa C is a 4 laser, 16-parameter instrument equipped with Violet, Blue, Yellow-Green and Red lasers, similar to the Fortessa X20 analysers in the lab.
- BD FACSymphony
The Symphony is an advanced analyser equipped with 5 high-powered lasers including a 60mW UV laser and upgraded detector arrays capable of detecting up to 30 parameters simultaneously.
- Helios – Mass Cytometer
The latest CyToF instrument from Fluidigm that is capable of measuring greater than 40 parameters simultaneously. The instrument combines aspects of traditional flow and mass spectrometry measuring mass instead of fluorescence.
- Miltenyi GentleMACS Octo Dissociator
This system is a benchtop instrument for semi-automated and standardised tissue dissociation or homogenisation of up to eight samples. Single-cell suspensions or thorough homogenates from virtually any tissues are easily and reproducibly obtained using the unique C Tubes or M Tubes. These single-use gentleMACS tubes allow sample preparation in a closed and sterile system, providing a high level of user safety and minimise cross-contamination.
- NovoCyte Quanteon
The latest 4 laser (Violet, Blue, Yellow-Green and Red) laser flow cytometer from Agilent, capable of measuring up to 25 parameters simultaneously. The Quanteon detectors are capable of measuring signals over a 7.2 log dynamic range, eliminating the need for voltage optimisation. The instrument can hold a 40-tube rack as well as 24, 48 or 96-well plates for high-throughput analysis of samples. Data analysis can be performed during sample acquisition such that all statistical tables and plots can be generated before the end of the run. Automated cleaning and shutdown also allow for users to walk away from the instrument during acquisition.
- CyTOF XT
The latest mass cytometer from Fluidigm capable of measuring over 50 parameters simultaneously. The instrument combines mass spectrometry with flow cytometry, measuring the time of flight of heavy metals to distinguish markers rather than fluorophores. The XT also contains a 13-sample, refrigerated carousel and autosampler, allowing users to run multiple samples in an automated manner.
Sorters
- BD FACS Aria Fusion
Aria Fusion is a 5 laser sorter capable of measuring 20 parameters including FSC and SSC, and can sort up to 4 distinct populations simultaneously, with different cell collection devices including tubes, slides, and 6/96/384 well plates. Thermal control is installed on the sorter to maintain sample at user-defined temperatures. The instrument is housed in a class 2 biosafety cabinet and is operated by members of the TTP.
- BD Aria III
Aria III-A is a 3-laser (Violet, Blue and Red) sorter capable of measuring 16 parameters.
Aria III-B is a 4-laser (Violet, Blue, Yellow-Green and Red) sorter capable of measuring 18 parameters.
Both sorters are able to sort up to 4 different populations simultaneously, with different cell collection devices including tubes, slides, and 6/96/384 well plates. Thermal control is installed on the sorter to maintain sample at user-defined temperatures. The instrument is housed in a class 2 biosafety cabinet and is operated by members of the TTP.
- MACSQuant Tyto
The Tyto cell sorter is a 3 laser (Violet, Blue and Red) benchtop sorter capable of measuring up to 8 parameters. It is user-operated and ensures full sterility through the use of fully closed MACSQuant Tyto cartridges, eliminating risk of contamination and aerosol generation. Setup is easy and does not require drop delay or laser alignment. The sorter is also gentle to cells due to the low pressure at which the cartridge operates.
Services
- Cell Sorting Service
The facility runs a cell sorting service with facility members of staff working closely with researchers to help develop effective sorting experiments.
- Mass Cytometry Service
The facility runs a mass cytometry service running samples for researchers in the institute and ensuring optimal performance of the instrument.
- Panel Design and Project Development
The facility provides expertise in panel design and project development helping researchers to get the most out of the instrumentation and the fluorescence/mass technology currently available.
- Instrument Training
Training is provided on the analysers to researchers of all levels with the aim of making the state of the art technology accessible to researchers with varying level of expertise.
- Software available
- FlowJo
- FACSDiva
- Cytobank
Resources
If you have any comments or queries regarding the CI Translational Technology Platforms please contact us at: ci.ttpfeedback@ucl.ac.uk or come along to one of our drop-in clinics.
