University of Leeds Faculty of Medicine and Health Flow Cytometry and Imaging Facility

Flow Cytometry and Metabolimetry Facility

What is flow cytometry?

Flow cytometry is an invaluable tool to provide high-end statistical sampling on large populations of cells in suspension. This technology allows for multiparameter assessments based on protein expression, content of cellular function dyes or specific nucleotide labels.

Fluorescent Activated Cell Sorting (FACS) can be utilised to purify target cell fractions for downstream work, including genomic sequencing and clonagenic assays.

Equipment

Flow Cytometry Analysers

The facility is equipped with 3 flow cytometry analysers and is in the process of sourcing a 4th analyser. Our systems have a range of capabilities including high throughput and high volume sampling and up to 18 simultaneous parameter collection on systems equipped with a plate arm. All our systems benefit from digital processing (Pulse Area, Height & Width measurements) and can be readily reserved thorough the QReserve online calendar system. Offline analysis of data can be carried out on any of our 4 offline data analysis PCs free of charge. All our flow cytometer analysers are user-operated following specific training for the instrumentation from facility support staff. To organise training on a system please email: medlfacs@leeds.ac.uk.

Attune (Thermofisher) – Room 5.04b WTBB

The Attune digital 2 laser 6 colour is an acoustic focusing analyser for improved small particle resolution and high speed/high volume sampling and is our fastest system. It features 3 parameters from a 488nm blue laser and 3 parameters from a 405nm violet laser (For details click here).

Cytoflex S (Beckman Coulter) – Room 5.04b WTBB

The Cytoflex LX is our newest digital 6 laser 21 colour flow cytometer analyser, offering improved small particle resolution and high speed/high volume sampling. Options for 2 SSC parameter detections from both the Violet 405nm laser and blue 488nm laser simultaneously for enhanced small particle detection for particles down to 200nm in size. PC is Windows 7 64bit with CytExpert V2.3 Software for data acquisition. This system is user run, but training is required from facility support staff prior to access being given (For details click here).

Cytoflex S (Beckman Coulter) – Room 5.04b WTBB

The Cytoflex S is a digital 4 laser 13 colour flow cytometer analyser for improved small particle resolution and high speed/high volume sampling. It features options for 2 SSC parameter detections of the Violet 405nm laser and blue 488nm laser simultaneously for enhanced small particle detection for particles down to 200nm in size (For details click here).

LSRII (BD Biosciences) – Room 5.04b WTBB

The LSRII is our optically most capable digital 4 laser 16 colour flow cytometer analyser. The 355nm UV Laser options allow for Ca2+ ion flux measurement with the Indo-1 label in calcium signalling assessments. The system is fitted with a HTS plate arm and is well suited to run cytokine bead based multiplex arrays (For details click here).

Fluorescence Activated Cell Sorting

The facility is currently equipped with two state-of-the-art high-speed cell sorters on the St James's University Hospital Campus UoL. All our systems are capable of sorting into FACS tubes and into a variety of collection plates with index sorting capability.

Influx (BD Biosciences) – Room 6.04a WTBB

The Influx is our high-end digital cell sorter housed in a CAT 2 hood allowing for the sorting of unscreened patient samples and other more high-risk samples including transiently transduced cell material (the facility will require a copy of the risk assessment of material prior to sorting). This system is run as a service by facility support staff due to the complexity of the 5 laser, 19 colour parameter system. Bookings must be made using the QReserve Online Calendar System. Please confirm staff and system availability and email medlfacs@leeds.ac.uk to confirm the sorter booking with staff (For details click here).

FACS Melody (BD Biosciences) – Room 6.32 CSB

The FACS Melody is our newest digital cell sorter. This easy to use 2 laser 6 colour system can be run by researchers (training is provided by support staff) or by facility support staff (dependent on staff availability through QReserve). For training requests or to arrange for staff to operate the cell sort please check staff and system availability and email medlfacs@leeds.ac.uk to confirm the sorter booking (For details click here).

Metabolimetry

Seahorse XFe96 (Agilent) – Room 6.04a WTBB

The Seahorse XFe96 is a plate-based fluorescent sampling system configured to assay cells in 96-well plates, allowing for maximal sampling of test criteria. This technology allows for the determination of mitcochondrial respiration and glycolysis through the measurement of the Extracellular Acidification Rate (ECAR) and the Oxygen Consumption Rate (OCR) on cells placed under defined stressed conditions (For details click here).

The facility offers training for this system and following training this system can be booked in using the QReserve online calendar system. We also provide a resource to purchase assay kits directly through the facility expediting the speed with which projects can be set up. To organise training on a system or to purchase Seahorse reagents please email: medlfacs@leeds.ac.uk.

Services

We offer our researchers and our external collaborators a range of services and support to allow a streamlined workflow from assay design through to publication. The core services offered by the FoMH core Flow Cytometry & Imaging Facility are:

Project Design
Grant Costings
Experimental Planning & Panel Design
Cell Purification Applications
Applications Workshops
Equipment Training
Teaching Applications
Data Analysis Support
Experimental Troubleshooting
Outputs for Data Publication

Facility Charges

Required Equipment and Facility Charges

Combined facility annual registration and access charge £50/yr
BD Biosciences Influx 6 Way Cell Sorter - £64.55/hr (plus 1hr Non-Sterile, 1.5hr Sterile Sort Setup Charge) staff run system
BD Biosciences FACS Melody 2 Way Cell Sorter - £55/hr (user run system plus £30 training charge)/£64.55/hr for staff operated service
BD Biosciences LSRII FACS Analyser 18 colour 4 laser with High-Throughput Plate Arm - £44/hr
Beckman Coulter Cytoflex LX FACS Analyser 21 colour 6 laser with High-Throughput Plate Arm - £42.89/hr
Beckman Coulter Cytoflex S FACS Analyser 13 colour 4 Laser with High-Throughput Plate Arm - £42.89/hr
Thermofisher Attune FACS Analyser 6 colour 2 laser - £33.39/hr
Agilent Seahorse XFE96 Cellular Metabolic Plate Analyser - £44.35/run
Agilent Seahorse XFE96 Reagents (email facility for pricing)
Additional staff support for data analysis or acquisition on top of training or setting of instrument settings charge at £30/hr