Flowcytometry Core Facility

Contact: Mr Andrew Worth, Flow Sorting Facility Manager
Tel: +44 (0)1865 617640
Email: andrew.worth@ndm.ox.ac.uk

Background

Flow cytometry allows the simultaneous measurement of certain characteristics of individual cells based on light signals generated when fluorescently labelled cells are made to flow past one or more laser beams. Prime examples of cell characteristics that can be analysed include size, granularity, phenotypic markers, intracellular proteins, DNA content and calcium flux. Flow sorting extends flow cytometry to the collection of subsets of a mixed cell population, in a purified form, by the electrostatic deflection of droplets containing target cells away from the main flow stream and into a collection tube.

The Flow Cytometry Lab has been set up as a Small Research Facility that is part of the Jenner Institute, in the Old Road Campus Research Building, Oxford.

Services provided

The facility provides two main flow cytometer based services: flow analysis and flow sorting. These services are available to all members of Oxford University, other academic institutions, and industry.

Training and/or experimental and technical assistance is provided as necessary. On-site data analysis may be performed with FlowJo, FACSDiva or Summit.

For flow analysis there are 2 digital cytometers and an old analogue cytometer available for booking. The user has the option of having the samples run for them, in which case they will need to liaise with the facility manager, or they can run their own samples after suitable training.

For cell sorting there is a single high-speed cell-sorter that has a dedicated operator to run the user’s samples. Up to 4 populations can be sorted simultaneously or there is the option to sort into multi-well plates. In all cases the user prepares their own samples.
 

Facility equipment

Becton Dickinson LSR II (purchased 2007)

15-colour digital flow analyser

• Three solid-state lasers (488nm, 635nm, 405 nm)
• Analysis-only instrument (up to 18,000 events/s)

• PC-based (WinXP Pro) digital compensation software (FACSDiva v6.1) for acquiring/analysing FCS 3.0 data
• Data Storage: hard drive and network storage areas
• Data may be analysed using third-party flow cytometry software, such as FlowJo

Available parameters (most signals may be defined as linear peak, log peak or area):

• Forward Scatter (FSC)
• Side Scatter (SSC)
• Fluorescent signals (FL1 – FL15), which may be used in various configurations, with up to 6 colours from the 488nm laser, 6 colours from the 405nm laser and 3 colours from the 635nm laser

Beckman Coulter CyAn™ ADP LX High-Performance Research Flow Cytometer (purchased 2005)

9-colour digital flow analyser

• Three solid-state lasers (488nm, 635nm, 405 nm)
• High-speed (up to 50,000 events per second) analysis-only instrument
• PC-based (WinXP Pro) digital compensation software (Summit 4.3) for acquiring/analysing FCS 3.0 data
• Data Storage: hard drive and network storage areas
• Data may be analysed using third-party flow cytometry software, such as FlowJo

Available parameters (most signals may be defined as linear peak, log peak or area):

• Forward Scatter (FSC)
• Side Scatter (SSC)
• Fluorescent signals (FL1 - FL9), which may be used in various configurations, with up to 5 colours from the 488nm laser, 2 colours from the 405nm laser and 2 colours from the 635nm laser
• Time and calculated parameters such as ratios

Beckman Coulter Legacy MoFlo MLS High Speed Cell Sorter

7-colour digital fluorescence activated high-speed cell sorter (purchased 2005)

• Must be run by dedicated operator
• Two lasers: Blue diode (488nm) 200mW max, Red diode (635nm) 35mW
Capable of sorting at rates up to 30,000 events/second
• Up to 4 subpopulations can be collected simultaneously
Fitted with CyClone unit for single-cell sorting into multi-well plates
• Temperature control of sample and collected populations available
• PC-based (WinXP Pro) digital compensation software (Summit 4.3) for acquiring/analysing FCS 3.0 data and setting sort decisions
• Data Storage: hard drive, and network
• Data may be analysed using third-party flow cytometry software, such as FlowJo

Available parameters (most signals may be defined as linear peak, log peak or area):

• Forward Scatter (FSC)
• Side Scatter (SSC)
• Fluorescent signals (FL1 - FL7), which may be used in various configurations, with up to 5 colours from the 488nm laser and 2 colours from the 635nm laser
• Time and calculated parameters such as ratios

Becton Dickinson FACSCalibur (purchased 1996)

4-colour analogue flow analyser

• Two solid-state lasers (488nm and 635nm)
• Analysis-only instrument ( • Mac-based (OS 9.2) analogue compensation software (CellQuest Pro) for acquiring/analysing FCS 2.0 data
• Data Storage: hard drive
• Data may be analysed using third-party flow cytometry software, such as FlowJo

Available parameters (most signals may be defined as linear peak or log peak):

• Forward Scatter (FSC)
• Side Scatter (SSC)
• Fluorescent signals (FL1 – FL4), which may be used in various configurations, with up to 3 colours from the 488nm laser and 1 colour from the 635nm laser

We welcome opportunities for collaboration or business partnership; enquiries can be directed to Mr Andrew Worth:
Email: andrew.worth@ndm.ox.ac.uk
Tel: +44 (0)1865 617640