Using multicolor flow cytometry to identify, characterize and isolate cell populations

Flow cytometry & FACS sorting shared facility

The department of Immunology has several state-of-the-art flow cytometry instruments available for users within the department but also outside users. These instruments are user-operated but support is available through the two sort operators and the manager. Furthermore, the department of Immunology is a member of the FACS Sorting Shared Facility (FSSF) and has two ARIA FACS sorting instrument, one of them situated in a class II cabinet, to serve the sorting needs of the department and outside users. The two dedicated sort operators Peter van Geel and Harm de Wit are performing the sorts for internal and external sorts. Yvonne Müller is the scientific manager of the flow cytometry facility.

Analysers

The following analyzers are available:

FACSCanto II (BD Biosciences)​

  • 3 lasers (Violet Laser – 405nm, Blue Laser – 488nm, Red Laser – 633nm)
  • 8 parameters

LSRFortessa SOP (BD Biosciences)

  • 4 lasers (Violet Laser – 405nm, Blue Laser – 488nm, Yellow/Green Laser – 561nm, Red Laser – 640nm)
  • 16 parameters

Aurora (Cytek)

  • Spectral flow cytometry instrument
  • 5 lasers (UV laser – 355nm, Violet Laser – 405nm, Blue Laser – 488nm, Yellow/Green Laser – 561nm, Red Laser – 640nm)
  • 64 detectors
  • 35+ parameter instrument

Instruments

The following sort instruments are available:

1. FACSAria III (BD Biosciences)

  • 4 laser (Violet Laser – 405nm, Blue Laser – 488nm, Yellow/Green Laser – 561nm, Red Laser – 633nm)
  • 15 parameters
  • Class II Cabinet

2. FACSAria II (BD Biosciences)

  • 3 lasers (Violet Laser – 405nm, Blue Laser – 488nm, Red Laser – 633nm)
  • 9 parameters

Additional, four FACSCanto II (BD Biosciences) and two FACSLyric (BD Biosciences) flow cytometry instruments are available for diagnostics, clinical trials and translational research within the Laboratory Medical Immunology (LMI) of the department of Immunology.

FACS Sorting Shared Facility (FSSF)

The department of Immunology together with the departments of Cell Biology, and Pathology established the FSSF of Erasmus MC to transfer knowledge, provide high quality sorting and technical service, and optimize usage of sort instruments. The FSSF serves members of Erasmus MC and outside academic and non-academic users. Human and animal primary cells and cell lines are sorted based on fluorescence probes (e.g. antibodies, fluorescent proteins, dyes). Five flow cytometry sort instruments (FACSAria II and III, BD Biosciences) with 4-5 lasers (UV laser – 375nm, Violet Laser – 405nm, Blue Laser – 488nm, Yellow/Green Laser – 561nm, Red Laser – 633nm) are available for bulk and single cell sorts of non-GMO and non-biohazard cells. ML-I and ML-II GMO material as well as BSL-1 and BSL-2 material can be sorted on the instrument situated in a class II cabinet (FACSAria III). Four sort operators are available for the sorting request of the users of the FSSF. The manager of the FSSF is Yvonne Müller.

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