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Flow cytometry facility

Flow cytometry is a laser based, biophysical technology used for cell counting, sorting, and biomarker detection. Cells are suspended in a stream of fluid and passed by an electronic detection apparatus. It allows simultaneous multi-parametric analysis of the physical and/or chemical characteristics of up to thousands of cells per second. Based on the characteristics cells can also be sorted with high precision allowing subsequent experiments with well-defined cell populations.

The Netherlands Cancer Institute has a Flow cytometry facility with several analyzers and two sorters. The two operators maintain the equipment, give advice and training and perform sorting experiments.

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 six flow cytometry analysers and two high speed cell sorters in the facility.

  • BD LSRII with 5 lasers (488 nm, 355 nm, 405 nm, 561 nm and 635 nm) and 18 fluorescence detectors
  • BD Fortessa with 4 lasers (488 nm, 405 nm, 561 nm and 635 nm) and 15 fluorescence detectors equipped with a HTS-loader for sampling out of a multiwell plate
  • Two  Becton Dickinson FACSCalibur analysers, one of which is capable of sampling out of a multiwell plate, with 2 lasers (488 nm, 635nm) and 4 fluorescence detectors
  • Beckman Coulter Cyan, 3 lasers (488nm, 405 nm,633 nm) and 9 fluorescence detectors
  • BD FacsArray, 2 lasers (532 nm and 635) and 4 fluorescence detectors capable of sampling out of a 96-wells plate
  • BD FacsAria Sorter with 3 lasers (488nm, 405 nm,633 nm) and 9 fluorescence detectors for high speed 4-way bulk sorting and single cell deposition sorting.
  • Beckman Coulter Moflo Astrios 4 lasers (488nm, 405 nm, 561 nm

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

Experience

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Martijn van Baalen

Flow cytometry operator

Experience

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