Flow cytometry core service

The Medical School's flow cytometry facility is a core facility based in the RHH Medical School and provides a service to all at the University and externally.



Flow cytometry is a technique that enables single whole particles/cells to be analysed. Events pass through lasers and physical properties such as relative size (FSC) and granularity/internal complexity (SSC) are detected. If there is any fluorescence also associated with the cell this can also be seen, with the emitted light being diverted via a series of mirrors and filters and collected by PMTs with specific bandpass filters in front of them.

Phycoerthrin (PE) - Excitation Max 561 nm

  • Emission Max 576nm 
  • Detection Band pass filter 575/26nm (ie 562-588nm)

Using physical or fluorescent properties, cells can be separated from a parent population into different tubes, this is called cell sorting. Cells can be bulk sorted into tubes, or single cells can be sorted into 96/384 well plates.

Example applications

  • Immunophenotyping (up to 40 colours on Cytek Aurora 5 laser)
  • Cell cycle
  • Viability/Apoptosis/Necrosis
  • Calcium flux
  • Cell counts
  • Detection of Extracellular vesicles, bacteria, algae
  • Transfection efficiency (e,g GFP, mCherry)

Services provided by the facility

The facility comprises

  • 2 Cytek Aurora spectral analysers
  • 1 LSRII analyser
  • 1 FACSMelody cell sorter
  • A soluble cytokine quantification service
  • Analysis software FCSExpress (available from August 2023)

Unlike the traditional flow cytometers which detect only a small portion of a fluorochromes spectral emission, the spectral analysers collect all the light emitted. This enables multiple fluorochromes to be separated and analysed simultaneously.

In May 2023 we took delivery of a second Cytek Aurora. This is a five-laser system with a plate loader capable of detecting up to 40 different colours.

Aurora spectral analyser

View spectrum examples

Our staff can help you with all aspects of your work including planning and running experiments and data analysis.

Bookings and charges

Technical time can be booked on the Flow Google Calendar and all the machines can be booked through the Flow Lab on Clustermarket. Please contact the facility for access.​​​​

Contact us at flow@sheffield.ac.uk.


LSRII: £45 per hour

Aurora 3-laser or 5-laser: £55 per hour


FACSMelody - £80

Filters and configurations


UV laser 355nm

UV 450/50

UV 530/30

Violet laser 405nm

Violet 450/40

Violet 525/50

Blue laser 488nm

Blue 530/30

Blue 575/26

Blue 610/20

Blue 660/20

Blue 695/40

Blue 780/60

Red laser 633nm

Red 660/20

Red 730/45

Red 780/60


Blue laser 488nm



Yellow/green 561nm





Red laser 640nm



Aurora (3-laser)

Violet 405nm

16 channels (420-829nm)

Blue 488nm

14 channels (498-829nm)

Red 633nm

8 channels (652-829nm)

Aurora (5-laser)

UV 355nm

10 channels 365-829nm

Violet 405nm

16 channels 420-829nm

Blue 488nm

14 channels 498-829nm

Yellow-Green 561nm

10 channels 567-829nm

Red 640nm

8 channels 652-829nm

Helpful websites


Cytek Biosciences

BD Biosciences


BD Biosciences


Antibody suppliers

Cytek Biosciences

Thermo Fisher Scientific


BD Biosciences Reagent Catalogue

A global reputation

Sheffield is a research university with a global reputation for excellence. We're a member of the Russell Group: one of the 24 leading UK universities for research and teaching.