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Flow cytometry instruments and configuration

Analysers

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

We have 4 analysers:

The Attune NxT is equipped with four lasers (488nm (blue), 637nm (red), 405nm (violet) and 561nm (yellow-green)) and is capable of detecting 16 parameters (FSC, SSC, and 14 fluorescent parameters). Equipped with an autosampler compatible with many different plate formats, including 96-well, 384-well, and deep-well plates. The Attune is equipped with acoustic-assisted hydrodynamic focusing and is able to achieve sample-throughput rates up to 10 times faster than traditional cytometers. This means that you can process samples more quickly and acquire sufficient events even from very dilute samples. In addition, it accommodates no-wash, no-lyse protocols, allowing you to run whole blood without lysis. More information about acoustic focusing can be found here: https://www.thermofisher.com/uk/en/home/life-science/cell-analysis/flow-cytometry/flow-cytometers/acoustic-focusing-technology-overview.html

The Attune Nxt configuration is the following:

Attune

 The CytoFLEX S is equipped with four lasers (488nm (blue), 640nm (red), 405nm (violet) and 561nm (yellow-green)) and is capable of detecting 15 parameters (FSC, SSC, and 13 fluorescent parameters). It can run samples from 96-well plates. The CytoFLEX S is designed to be user-friendly and users should be able to run it independently after a training session with a member of the facility staff. For more information about the Cytoflex S please visit https://www.mybeckman.uk/flow-cytometry/instruments/cytoflex-s

The cytoFLEX S configuration is the following:

 cytoflez

 

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. 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 5 lasers at 488nm (blue), 561nm (yellow-green), 633nm (red), 355nm (UV) 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.

aurora5L

 


Cell sorters

We currently have 3 cell sorters: the BeckmanCoulter MoFlo Astrios, the BD Aria III 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. 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 III has 4 lasers (405, 488, 561 and 633nm), 16 fluorescent detectors and 2 scatters. The Aria III is also contained within a class II biohazardous cabinet and can be used to sort CLASS I and II samples. The Aria can also sort cells in bulk (up to 4 populations simultaneously) or into multiwell trays or slides. The Aria III standard configuration is the following: 

Aria III

 

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 for the University of Cambridge users. Please read the Flow cytometry charges section for more information. 



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 FlowJo. Other available software: Kaluza, FCS Express 6, Summit, Attune NxT software, SpectroFlo, CytExpert.