Flow Cytometry

Introduction :

The flow cytometry platform of the IMRB is located on the 1st floor of the Research building of the Henri Mondor hospital.

It is equipped with two cell sorters (BD Influx Sorp and BD Aria Fusion) and two analysers (BD Fortessa X20 and Miltenyi Biotec MACSQuant 16), allowing to characterize and isolate elements (cells, bacteria, …) from a mono-dispersed suspension specifically fluoresced.

The projects developed on the platform are of various origins but mainly concern research in immunology, including the study of immune function, drug resistance, cell proliferation, apoptosis and the cell cycle.

 

Competence and Expertise :

  • Scientific and technical fields: cell biology, immunology
  • Competencies and know-how: Flow cytometry-based cell analyses and sortings

Available Material :

The platform manages and ensures the maintenance and quality control of the following instruments:

 

 

  • INFLUX_pl_triCell sorter : Influx (BD Biosciences)

– 4 lasers (488, 640, 405 et 561nm)
– 2 morphological parameters (size and density),
– 18 fluorescence parameters :
– Laser 488nm : 520/15, 530/40, 670/30 et 710/50.
– Laser 640nm : 670/30, 720/40 et 750LP.
– Laser 405nm : 460/50, 520/35, 610/20, 660/20, 710/50 et 750LP.
– Laser 561nm : 585/29, 610/20, 670/30, 710/50 et 750LP.
– Detection of microparticles (<0,2µm).
– 70 000 events per second.
– Up to 6 channels for sorting, 4 nozzle sizes (70, 86, 100 et 140µm).

– Monitoring of the temperature (5°C-95°C)

– Collect of purified cells in microtubes, 5 ml, 15 mL or 50 mL tubes. Possibility of cell cloning in micro-plates (6/24/48/96/384 wells) or on microscopic slides.

 

 

  • Cell sorter : Aria Fusion (BD Biosciences)

– 4 lasers (488, 640, 405 and 561nm)
– 2 morphology parameters (granulosity size)
– 16 fluorescence parameters
– Laser 488nm: 530/30, 695/40.
– Laser 640nm: 670/30, 730/45 and 780/60.
– Laser 405nm: 450/50, 525/50, 610/20, 660/20, 710/50 and 780/60.

– Laser 561nm:  582/15, 610/20, 670/14, 710/50, 780/60.
– Up to 4 sorting channels
– 3 types of nozzles (70, 85, 100µm)
– Recovery in FACS and eppendorf tubes or 15mL
– Culture plate cloning option
– Class II Type 2 Containment Enclosure

 

 

 

 

 

 

  • Analyser: LSRFortessa X20 (BD Biosciences)          

– 4 lasers (488, 640, 405 et 561nm)

– 2 morphological parameters (size and granulosity),

– 16 fluorescence parameters,

– Laser 488nm: 520/30, 695/40.

– Laser 640nm: 670/14, 730/45 and 780/60.

– Laser 405nm: 450/50, 525/50, 610/20, 670/30, 710/50 et 780/60.

– Laser 561nm: 586/15, 610/20, 670/30, 710/50 and 780/60.

– 20 000 events per second.

 

 

 

 

 

 

 

 

  • Analyser: MACSQuant 16 (Milteny Biotec)
    – 3 lasers (405, 488 and 640nm)
    – 2 morphology parameters (size and granulosity)
    – 14 fluorescence parameters
    – For 488nm laser: 525/50, 579/34, 615/20, 667/30, 725/40 and 785/62.
    – For 640nm laser: 667/30, 725/40 and 785/62.
    – For 405nm laser: 450/50, 525/50, 579/34, 615/20 and 667/30.
    – 15,000 events per second
    – Absolute numbering
    – Tube autosampler FACS, eppendorf, Falcon or plate
    – Resuspension of the sample (by suction)

 

 

 

 

 

 

 

 

  • Low-Jo analysis station

 

Proposed services :

  • Cellular sorting.
  • Flow cytometry analysis.
  • Training in the acquisition, analysis and interpretation of the results obtained.
  • Provision of cytometers (LSR-X20, MQ16 or Aria) after validation of the training.
  • Data analysis (Flow-Jo software).
  • Advice to users.

Conditions of Use :

The facility is opened to all members of IMRB as well as to academic and non-academic teams, provided they obtained agreement from the staff. People and teams willing to use the facility equipment must first meet and discuss with the facility staff in order to establish provisional planning.

 

Opening hours :

From Monday till Friday from 8h30 am to 5h30 pm. Reservation can be made by email at the following address: cytometrie.imrb@inserm.fr. Outside of the opening hours, getting access to the analyzers only requires agreement of the staff.

 

Location :

1st floor of the Research Building of the Henri Mondor Hospital

Doors AA-061 and AA-064

 

Contact :

Contact us

 

The documents for the agents provided by the cytometry platform can be found in the data library on the platform homepage.

 

 

 

 

ISO 9001 certification defines a set of requirements concerning the implementation of a quality management system, regardless of size and industrial activity.

 

 

 

 

 

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