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Flow cytometry facility

Flow cytometry is a powerful tool to quickly characterize millions of individual cells and, if needed, isolate the cells based on their characteristics. Cell identification is based on laser light scatter signals due to differential cell surface properties and intracellular components of the cell, and in combination with fluorescent signals from antibody-fluorochrome conjugates, fluorescent proteins and other fluorescent dyes that may be used to tag intracellular or extracellular markers.

The flow cytometry facility gives researchers access to a variety of analytical flow cytometers and cell sorters. Facility staff maintains a high-standard quality assurance for all equipment; provides professional advice on experimental and panel design for both analysis and cell isolation purposes; provides introductory and advanced training courses for independent use of the analytical flow cytometers and cell sorters.

 

Activities and services

Researchers are offered hands-on training to learn the principles of flow cytometry and to operate the analysers themselves. The analysers are available to trained users 24/7.
The ability to measure multiple parameters on individual cells is the most powerful aspect of flow cytometry. This allows flow cytometry to be used for a wide range of applications. Among the most common applications at the Netherlands Cancer Insitute are immune phenotyping with up to 18 colours, apoptosis assays using annexin V, cell cycle analysis, and analysis of cells expressing one or more fluorescent proteins. Lots of other applications are used and new applications are set up by the operators together with the researchers.

Through close cooperation of fluidics, laser technology and electronics it is possible to separate specific cells out of a cell suspension using the sorters at the facility. We are capable of sorting up to 16 colours into 6 fractions or into multiwall plates. We can also sorting into microwell slides. The development of these cells was followed under a confocal microscope at the digital microscopy facility.

 

Equipment

There are currently seven analytical flow cytometers and three high speed cell sorters in the facility.

Analytical flow cytometers

  • FACSSymphony A5 SORP (BD BioSciences): 5 lasers (488nm, 355nm, 405nm, 561nm and 638nm) and 28 fluorescence detectors.

  • LSRII SORP (BD BioSciences): 5 lasers (488nm, 355nm, 405nm, 561nm and 638nm) and 18 fluorescence detectors.

  • LSRFortessa SORP 2 (BD BioSciences): 4 lasers (488nm, 405nm, 561nm and 638nm) and 16 fluorescence detectors.

  • LSRFortessa SORP 1 (BD BioSciences): 4 lasers (488nm, 405nm, 561nm and 638nm) and 16 fluorescence detectors. Equipped with HTS-loader for read-outs from multi-well plates.

  • Attune NxT Accoustic flow cytometer (Invitrogen, ThermoFisher Scientific): 3 lasers (488nm, 405nm, and 638nm) and 11 fluorescence detectors. Equipped with autosampler for read-outs from multi-well plates.

  • FACSCalibur (BD BioSciences): 2 lasers (488nm, 635nm) and 4 fluorescence detectors. Equipped with HTS-loader for read-outs from multi-well plates.

  • FACSArray (BD BioSciences), 2 lasers (532nm and 638) and 4 fluorescence detectors capable of sampling out of a 96-wells plate.

Cell sorters

  • FACSAria Fusion SORP  (BD BioSciences): cuvette-based; 4 lasers (488nm, 405nm, 561nm and 638nm) and 16 fluorescence detectors. Capable of 4-way sorting in bulk in a variety of collection tubes and / or single cell index sorting into multi-well plates.

  • Moflo Astrios (Beckman Coulter): jet-in-air; 4 lasers (488nm, 405nm, 561nm, 638nm) and 16 fluorescence detectors. Capable of 6-way sorting in bulk in a variety of collection tubes and / or index sorting into multi-well plates.

  • FACSAria IIu (BD BioSciences): cuvette-based; 3 lasers (488nm, 405nm, 638nm) and 9 fluorescence detectors for high speed 4-way bulk sorting and single cell deposition sorting.

People working at the the Flow cytometry facility

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Anita Pfauth

Flow cytometry operator

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Frank van Diepen

Flow cytometry operator

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Martijn van Baalen

Head Flow cytometry

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Debajit Bhowmick

Flow cytometry operator

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