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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 illuminated characteristics of individual cells depending on light signals generated when fluorescently labelled cells are made to flow across one or more laser beams. Prime examples of cell characteristics that can be detected include size, granularity, phenotypic markers, intracellular proteins, DNA content and calcium flux.

Flow sorting extends flow cytometry to the detection and collection of cell subsets from mixed cell populations by the electrostatic deflection of droplets containing target cells away from the main flow stream into a suitable collection receptacle.

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 (ORCRB), Oxford.

Services provided

quantum-dots.jpeg

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 or FACSDiva.

For flow analysis there are 2 digital cytometers 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)

lsrii.jpeg15-colour digital flow analyser
  • Three solid-state lasers (488nm, 635nm, 405 nm)
  • Analysis-only instrument (up to 18,000 events/s)
  • PC-based (Win10) digital compensation software (FACSDiva v9.0) 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

BECTON DICKINSON LSR Fortessa X20 (PURCHASED 2018)

Jenner_FortessaX20.jpg18-colour digital flow analyser

  • Five solid-state lasers (488nm, 635nm, 405 nm, 561nm and 355nm)
  • Analysis-only instrument (up to 18,000 events/s)
  • PC-based (Win 10) digital compensation software (FACSDiva v9.0) 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 – FL18), which may be used in various configurations, with up to 2 colours from the 488nm laser, 5 colours from the 561nm laser 6 colours from the 405nm laser, 3 colours from the 640nm laser and 2 colours from the 355nm laser
  • Time and calculated parameters such as ratios

BECTON DICKINSON FACSARIA FUSION WITH BIOLOGICAL SAFETY HOOD (PURCHASED 2020)

Jenner_Fusion.jpg18-colour digital fluorescence activated high-speed cell sorter 

  • Must be run by dedicated operator
  • Five lasers: Blue (488nm) 50mW max, Red (640nm) 100mW, Violet (405nm) 85mW max, Yellow-Green (561nm) 50mW max and UV (355nm) 15mW max. Capable of sorting at rates up to 30,000 events/second
  • Up to 4 subpopulations can be collected simultaneously. Fitted with ACDU for single-cell sorting into multi-well plates
  • Temperature control of sample and collected populations available
  • Equipped with Class II Type A2 biosafety cabinet
  • PC-based (Win10 Pro) digital compensation software (FACSDiva v9.0) 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 – FL18), which may be used in various configurations, with up to 2 colours from the 488nm laser, 3 colours from the 640nm laser, 6 colours from the 405nm laser, 5 colours from the 561nm laser and 2 colours from the 355nm laser
  • Time and calculated parameters such as ratios

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