FACS Sorting Shared Facility (FSSF)

Sorting cell populations based on fluorescence probes (e.g. antibodies, fluorescent proteins, fluorescent dyes)

• Human and animal cells (primary cells, cell lines)
• ML-I and ML-II GMO material as well as BSL-1 and BSL-2 material
• Bulk sorts
• Single cell sorts

Cell sorting instruments (specifications sheet):

• FACSAria III: 4 lasers, 17 parameters; 
  this sorter is situated within a class II biosafety cabinet  which allows for ML-I and -II as well as BSL-1 and -2 sorts;
• FACSAria II SORP: 5 lasers, 20 parameters
• FACSAria III: 4 lasers, 15 parameters
• FACSAria III: 4 lasers, 16 parameters
• FACSFusion: 5 lasers, 20 parameters; 
  this sorter is situated within a class II biosafety cabinet  which allows for ML-I and -II as well as BSL-1 and -2 sorts

Cell sorting examples

• GFP-expressing cells from a GFP-transfected cell lines;
• Naïve B cells from a buffy coat
• Virus-specific CD8+ T cells from a murine spleen
• Naïve CD4+ and CD8+ T cells from cord blood

The FSSF serves members of Erasmus MC and outside academic and non-academic users.

1. Sorting Fees

• Users from the departments of Immunology, Cell Biology and Pathology: please contact the manager or the operators
• Users from other departments within Erasmus MC: € 80/hour
• Academic users outside of Erasmus MC: €110/hour
• Non-academic users: €140/hour

2. Training Fees

• Advanced training for sort operators: €500/trainee

  (3 observation sort sessions, 2 hands-on sessions, 3 months of advice)

• Multicolor flow cytometry (LSR Fortessa): €200/trainee (introduction to flow cytometry theory, instrument introduction, 1 hands-on session)

3. Advice

• €40/hour(in person or by phone/email from an operator)

These fees cover part of the expenses for personal, instrument service contracts and consumables.

The FSSF supports users of the sort instruments as well as other flow cytometry analysis instruments with:

• Experimental design
• Sorting strategies
• Optimize protocols for cell sortingpreparation

• For sorting availabiliy:
  check the reservation calendar from the online booking system
  
  
• Please provide us your experiment specifications (below) in a email or document and send it to: cellsorting@erasmusmc.nl
  

  Experiment specifications (set-up):

  • Your name:
  • Your emailaddress:
  • Your telephone number:
    
    
  • Details about the requested sort: 
      date:
      time:
      preference for department / type of instrument / sort operator:
    
  • Does GMO of biosafety apply? GMO / biosafety / no
    • If yes to GMO:
      • please provide the license number:
      • Is this license number already approved for sorting: yes / no
    • If yes to biosafety:
      • What kind of biosafety: human cells / infections / other biosafety?
      • If it concerns 'other biosafety', what kind of biosafety?
      • Is this sort of biosafety material approved by BVF (Biologisch Veiligheidsfunctionaris) yes /no
          
  • Please indicate the following required information:
    • What kind of cells?
    • How many cells will be sorted?
    • What is the percentage of the population to be sorted?
    • How many sorted cells are necessary?
    • Any other important information:
  
  
  
• The facility will contact you for confirmation 
  (and will reserve a timeslot, this is only possible when complete information is provided)

For all other questions use our webform below this website

• Advanced training for sort operators including instrument set up, quality control and instrument maintenance;
• Multicolor flow cytometry including instrument and experimental set up, background of flow cytometry

The FACS Sorting Shared Facility (FSSF) was set up in 2015 through an initiative from the department of Immunology and the chairs and departmental members involved in flow cytometry of Immunology, Cell Biology and Pathology. The goal is to support the Erasmus MC community with their sorting needs, transfer flow cytometry knowledge, improve the quality of sorts and optimize the usage of the sort instruments available within these three departments. Other departments are invited to also join this shared facility with their sort instruments.

Flow cytometry is a laser based technology to examine the cellular expression of antigens, DNA and other molecules using fluorochrome-conjugated molecules like antibodies. Single cell suspensions of a cell population stained with fluorochrome-conjugated molecules are intercepted by lasers , the laser excites the fluorochromes and the emitted light signal is visualized by a software. Using defined antigen expression, cells (and other cellular organisms like bacteria) can be sorted to high purity from a heterogeneous cell population using a cell sorter (Fluorescence-activated cell sorting, FACS).

Director of FSSF: Jan Dirk van de Hoef, Thema director D&A

Scientific manager: Yvonne Müller, Ph.D., Assistant Professor, department of Immunology; y.muller@erasmusmc.nl

Sort operators:

• Harm de Wit (department of Immunology)
• Peter van Geel (department of Immunology)
• Tsung Wai Kan (department of Cell Biology)
• Andrea Sacchetti (department of Pathology)

For questions please contact us via our contact email: cellsorting@erasmusmc.nl

The FSSF provides to service members of the Erasmus MC and outside users

• Fluorescence-based sorting (single cell sort, bulk sort)
• Setting up sort experiments
• Technical support for sort operators
• Supporting set-up of multi-parameter flow cytometry experiments

The FSSF performs sorts for

• member departments (Immunology, Cell Biology and Pathology)
• non-member departments including virology, rheumatology and oncology
• outside academic users include UMC Utrecht