Flow Cytometry

Flow Facility Equipment

All equipment is available for external use, please contact flow@babraham.ac.uk for more information about core facility and using the facility. Details of lasers and filters can be found by clicking on the instrument names.

Sorters

All of the sorters in the BI Flow Cytometry Core Facility are available as part of the cell sorting service run by the facility.

Flow kit

BD Aria III

4 lasers (488nm, 405nm, 561nm, 640nm) 16 fluorescent colours, capable of sorting 4 populations simultaneously. This machine can be used for plate sorting and index sorting.

BD Influx

5 lasers (488nm, 405nm, 561nm, 640nm, 355nm, 15 colours. Can separate 6 populations simultaneously. The Influx is in a Class 2 hood which allows sorting of human cell lines and human primary cells/tissues. This machine can be used for plate sorting and Index sorting.

BD FACS Aria Fusion

5 lasers (488nm, 405nm, 561nm, 640nm, 355nm, 18 colours. The Fusion is in a Class 2 hood which allows sorting of human cell lines and human primary cells/tissues. This machine, funded by the BBSRC, can be used for plate sorting and index sorting

Jazz

 

BD FACS Jazz x 2

3 lasers (488nm, 561nm, 640nm), 6 colours. The Jazz sorters are high-speed jet-in-air sorters that are ideal for sorting large numbers of cells, for example for CRISPR screening. They are also well-suited to sorting of smaller panels for example for fluorescent protein experiments. The Jazz sorters are capable of sorting 2 populations simultaneously. The machines can also do plate sorting and index sorting. Both Jazz sorters are housed in biosafety cabinets for containment level 2 work.

Bigfoot

 

Thermo Fisher Bigfoot  x3

The Bigfoot is a spectral cell sorter. It can carry out complex, high parameter analysis and sorting. The Bigfoot has 9 lasers (340nm, 405nm, 445nm, 488nm, 532nm, 561nm, 594nm, 640nm and 785nm) and 55 fluorescence parameters. It is capable of carrying out spectral and conventional flow cytometry. The Bigfoot can bulk sort at comparable speeds to other jet-in-air sorters (for example the BD Influx) and can sort up to 6 populations simultaneously. Plate sorting on the Bigfoot is incredibly quick due to the capability to sort multiple streams into a plate. A 96-well plate can be sorted in a few seconds. Other capabilities of the machine include straight-down sorting into plates, bulk sorting into multiple types of collection tubes at the same time, and sorting into 10x plates.

Analysers

Aurora analyser

Cytek Aurora Spectral Analyser

The Cytek Aurora measures the emitted fluorescence of dyes across the whole spectra using a range of detectors. The signal is stitched together to create a full spectrum signature for each fluorophore. Gathering data from all detectors allows the Aurora to resolve dyes that conventional cytometers cannot, so allowing a greater number of fluorochromes to be used in combination. The Aurora has 5 lasers, 3 scatter channels and 64 fluorescence channels. This allows complex, high parameter analysis to be carried out. Flat-top beam shaping allows for high flow rates without compromising on data resolution and there is also a module for analysis of samples from 96-well plates.

Fortessa machine

Fortessa 5a

This state of the art cytometer was purchased in April 2014, due to the popularity of our original Fortessa. With the increasing trend for polychromatic flow cytometry, this sophisticated equipment permits use of up to 18 fluorochromes simultaneously plus 2 scatter parameters.

This machine is proving an invaluable asset as it allows scientists to maximise data acquired from each sample, saving time, money and valuable cells/tissues. This machine has also been equipped with a UV laser, allowing scientists to use the new BD Brilliant UV dyes.

This Fortessa is also equipped with a High-throughput sampler, which can run 96 and 384 well plates.

 

Fortessa

Fortessa 5b

This analyser was added to the core in early 2012 (and upgraded to 5 lasers in 2016) and is a heavily used vital piece of equipment for the research that is carried out at Babraham Institute.

This machine allows 18 colours to be simultaneously measured as well as 2 scatter parameters. This machine is equipped with a 561nm laser, which allows detection of red fluorescent proteins and better excitation of PE fluorochromes. Utilising this laser line also reduces spillover and compensation with other channels such as FITC.

Since purchase this machine has proved very popular and is our most regularly used analyser.

Fortessa

Fortessa 5c

Our latest addition to the Flow Core was purchased in March 2016 to complement the exisiting Fortessa analysers.

As well as having 5 lasers for 18 colour analysis this instrument was configured to maximize the use of BD Brilliant UV dyes. It has been set up to allow 3 parameters to be analysed from the UV laser).

The new Fortessa is also equipped with a High-throughput sampler, which can run 96 and 384 well plates.

LSRII

BD LSRII

4 lasers (488nm, 355nm, UV, 405nm, 635nm), able to detect 10 colours.

The LSRII has been part of the core facility since 2006 and is a much-loved machine.

Equipped with a UV laser, it allows scientists to look at Calcium Flux, DAPI/Hoechst cell cycle as well as UV excited fluorochromes. 

Yeti

Yeti

Propel Labs YETI analyser allows users to take their analysis to another dimension. Not only is it a 5 laser, 30 parameter machine but it can be used for high-throughput analysis using tubes or plates. This machine allows simultaneous detection of 27 fluorescent and 3 scatter parameters (FSC and SSC from 488nm laser plus FSC from 405nm laser).

The filter configuration enables users to utilise a range of commercially available fluorochromes, allowing multi-colour phenotyping above the Fortessa’s capabilities whilst also giving the user more flexibility of dye combinations. The machine can be used in ‘stat’ mode to run individual tubes, but can easily be used with racks of 5ml and Eppendorf tubes to allow easy of use and walk away analysis.

Analysis of 96 well plates can be done in around 15 minutes making it perfect not only for in depth phenotyping studies but also screening. There is also a 384 well plate capability.

Flow lasers wide

Amnis Imagestream

4 laser (488nm, 405nm, 561nm, 640nm). This imaging cytometer produces multiple high resolution images of every cell directly in flow, giving quantitative information about each cell in up to 12 channels simultaneously.

The Imagestream is an image cytometer that allows quantitative data to be acquired as well as images of every event passing through the instrument to be recorded. This machine can analyse up to 5000 cells/sec. This clever machine allows the user to look at features of cells such as co-localisation, staining location, cell-cell location, spot counting and cell death analysis.

Members of the core facility are available to help set up and optimise assays and help with post-acquisition analysis of the data.