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Flow cytometry instruments and configuration

Analysers

We have 2 analysers.

The Cytek DxP8 provides 3 colour excitation at 488nm (blue), 561nm (yellow-green) and 637nm (red) with a maximum of 8 fluorescence and 2 scatter parameters. This cytometer is useful for anyone using one of the many flavours of red fluorescent protein (mRFP1, mCherry, dTomato etc.). The Cytek is based on FACScan fluidics and is designed to be user-friendly and we expect most users will be comfortable with it after one training session from the facility staff. The instrument configuration is the following:

Cytek DxP8

 

The Cytek Aurora is a different kind of flow cytometer which works by measuring the full spectral output excited by each laser and deconvoluting signals according to prerecorded fluorochrome characteristics. This means it is easier for you to run many different fluorescent probes, including ones previously too similar to use in combination (though they still need to be different spectrally to some degree). The Aurora has 4 lasers at 488nm (blue), 561nm (yellow-green), 633nm (red) and 405nm (violet). The Aurora has also a plate reader capable of running samples from 96-well plates. More information about the spectral analysis can be found at https://cytekbio.com/pages/aurora#tab-overview.

 

The analysers are user-operated but all users require at least one training session before they can be used independently. Training sessions should be requested on PPMS. 

 


Cell sorters

We currently have 2 cell sorters: the BeckmanCoulter MoFlo Astrios and the BD Aria IIu. 

The MoFlo Astrios has 5 lasers (355, 405, 488, 561 and 640nm) and a maximum of 18 fluorescent detectors and six scatter detectors. Most application will not use many of these data channels. It also is contained within a Class II Biohazard cabinet and in a designated Class II suite (with stand alone hood, centrifuge and incubator). We welcome enquiries for sorting Class II material (subject to risk assessment) but also expect to sort typical class I material. It can sort cells in bulk or deposit any chosen number in multiwell trays of up to 384-wells or onto slides. It takes about 2 minutes to 'clone' cells into a 96-well tray (assuming the desired population is about 1% of the input mixture). The machine can sort 6 different populations simultaneously, maximising the use of your samples. The Astrios standard configuration is the following:

 

The Aria IIu has 3 lasers (405, 488 and 633nm), 9 fluorescent detectors and 2 scatters. The Aria is not contained in a biohazard cabinet and can only be used with CLASS I samples. The Aria can also sort cells in bulk (up to 4 populations simultaneously) or into multiwell trays or slides.  

 

 We welcome  regarding possible uses.

Charges

In order to cover the running costs of the machines, we do have to make a charge for use. The analysers are charged at £23/hour of use and the cell sorters are charged at £58/hour.



Data Analysis


We also provide, free of charge, 2 stand-alone computer workstations for flow data analysis for users of the facility.  Both PCs run Summit and FlowJo.