Flow Cytometry

Flow cyotmetry facilityThe work horses of the Flow Cytometry Facility are 5 Cyan ADP 9 colour instruments and the recently acquired BD LSRFortessa X20

These instruments have been distributed amongst the Institute for Biomedical Sciences (IBR) and The Centre for Translational Inflammation Research (CTIR) making them locally available for many researchers. All the flow cytometers are looked after by Roger Birdwho will provide detailed training for anyone wanting to use these instruments. Once trained, you’ll be able to book the FACS analysers using Stratocore. Roger is also the key operator of the Moflo high speed cell sorter. Appointments for time on the sorter can be made thorough Roger who will advise on sample preparation, sorting parameters and will be present to operate the Moflo during the sort.

BD LSR Fortessa X20...

The Fortessa can analyse up to 16 colours (2 off the 488nm Laser, 6 off the 405nm Laser, 3 off the 635 Laser and 5 off the 560nm Laser) plus forward and Side Laser Scatter.

Cyan FACS Analyser...

Cyan ADP
The Cyans are capable of analysing up to 9 colours (5 off the 488nm Laser, 2 off the 405nm Laser and 2 off the 635 Laser) plus forward and Side Laser Scatter.

Moflo Legacy High Speed Cell Sorter...

The MoFlow cell sorter is operated by Roger Bird who you should contact to book time on this instrument.

It is equipped with three lasers and can detect up to 11 parameters. It is routinely used for high speed sorting with up to 50,000 cells per second, single cell sorting into PCR plates for gene expression analysis bottom right, or cloning.

Capabilities-

1 Sort 4 selected populations simultaneously

2 Enrich cell populations at rates of up to 80000 cells/sec

3 Purity sorts at rates of up to 30000 cells/sec

4 Sort cells directly into multi-well plates

Under the right conditions purity levels of >99 per cent and recoveries of >95 per cent can be achieved

lasers- 360,488,and 633nm

11 analysis channels- Forward scatter, side scatter,

5 fluorescence channels activated by 488nm laser

2 fluorescence channels activated by 360nm laser

2 fluorescence channels activated by 633nm laser

Suitable fluorophoresinclude FITC, RPE, PE-Cy5, PE-Cy7, PE-Texas Red, Hoechst, APC and APC-Cy7

FlowJo FACS Analysis Software...

The Technology Hub has a 25 seat license for the FlowJo FACS analysis software. The licenses are distributed throughout the College of Medical and Dental Sciences making it easily available to all FACS users. A full description of FlowJo and tutorials etc can be found at http://www.flowjo.com/learn/ .

MoFlo Astrios Flow Cytometer Cell Sorter...

The MoFlow Astrios is situated within the CTIR in the QE hospital

It is equipped with five lasers and can detect up to 18 parameters. Only sorter capable of >70,000 sort decisions/second validated performance with a variety of collection options including 6 way sorting and single cell sorting into PCR plates for gene expression analysis.

Capabilities and features-

Sort up to 6 selected populations simultaneously with a variety of temperature controlled (-4 to 40°C) collection options.

Plate sorting up to 1536-well

Mixed Mode sorting: each sort stream is capable of having its own sort mode (Enrich, Purify, Single) programmed. Soft aborts and waste can be collected for reanalysis or resorting.

Dual eFSC channels allow you to apply different application-specific masks for different particle types, or use different neutral density filters to allow a greater overall particle size range to be measurable simultaneously.

Contained within an NSF-49 tested Class II Biosafety Cabinet for greater sterility and biosafety.

lasers- 355, 405, 488, 561 and 642nm.

18 analysis channels- Forward scatter, side scatter,

3 fluorescence channels activated by 355nm laser

3 fluorescence channels activated by 405nm laser

6 fluorescence channels activated by 488nm laser

3 fluorescence channels activated by 561nm laser

3 fluorescence channels activated by 642nm laser

Suitable fluorophores include DAPI, Pacific Blue, FITC, PE, PE-Cy5, PE-Cy7, PE-Texas Red, APC and APC-Cy7