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Flow Cytometry Unit (FCU) Infrastructure for high-end FACS analysis and state-of-the-art cell sorting capabilities. Facility
Infrastructure for high-end FACS analysis and state-of-the-art cell sorting capabilities.
The FCU offers access to high-end FACS analysis and state-of-the-art cell sorting capabilities and actively provides scientists from abroad as well as from the University Medical Center Groningen with adaptive setups and dedicated support. The facility also provides services that include training, technical support, and assistance with experimental design and data analysis.

Actively providing scientists from abroad and the UMCG with adaptive setups and dedicated support. The facility also provides services that include training, technical support and assistance with experimental design and data analysis.

Booking information

Before you can use any instrument, you are required to:

  • Register as a facility user;
  • Follow individual training;
  • Create a personal account in Booked Scheduler;
  • Read all FCU rules and fee calculations.

    • Please complete and submit the online registration form.
    • Make sure that you upload a digital ML-II e-learning certificate (pdf, jpg, png or bmp), showing your name and the score obtained.

    • After registration, new users will receive an email with an appointment for a training session by one of the operators of the requested flow cytometer.
    • For the MoFlo Astrios and MoFlo XDP cell sorters, an introduction is not necessary as these systems are always controlled by the facility operators.

  • The facility uses a booking system for the reservations of the different flow cytometers. All users need to create an account.

    Once the introduction has been successfully completed, you will be allowed access to the flow cytometer by using the Booked Scheduler agenda.

  • The following rules and regulations apply to the people working at the FCU (ML-II classified):

    • Access to the FCU is restricted to those who are aware of the risks associated with entering this facility.
    • Work is not allowed to be started until the ML-II e-learning module has been completed and instructions have been given by the Responsible Staff Member (RSM) or another member of staff authorized by the biosafety officer (BSO) (e.g. the SM).
    • Maintenance staff must consult the RSM or SM before entering the room. All persons working with GMOs at level II must be registered with, and have the approval of, the BSO. This also applies to students and guest staff.
    • All persons working in an ML-II area must comply with any additional rules and measures and with any additional advice given by the BSO, RSM or SM.
    • In all cases not covered by these rules and regulations, the RSM, SM or BSO should be consulted.
    • Every lab worker is required to inform the RSM, SM or BSO in the event of irregularities, incidents, or emergencies.
    • For experiments with radionuclides, permission is also required from the Radiation Protection Unit.
    • The ML-II work instructions apply to all activities involving non-GMOs or GMOs classified as ML-I that are carried out in the ML-II room.
    • If work involving microorganisms classified as ML-III has to be carried out in the ML-II laboratory, permission must be obtained from the BSO.
    • Work involving GMOs must be specified in the GMO permit. The GMO permit contains additional instructions that must be followed.
    • Persons who have a digestive disorder or an open wound or are taking immunosuppressive medication may not be allowed to carry out certain work (please consult the BSO).

  • Your time slots in the Booked Scheduler will be used for fee calculations. The invoice will be sent to the responsible Principal Investigator (PI) at the end of each year.

    The invoice for internal users will be calculated as follows:

    • FacsVerse; Quanteon: €5/hr
    • LSR-II; FACS Symphony A50; Cytek Aurora: €10/hr
    • MoFlo XDP; MoFlo Astrios: €30/hr
    • Sony SH800s: 20 €/hr

    Users from outside the University Medical Center Groningen/University of Groningen:

    • Analysers: €200/hr
    • Sorters: €300/hr

A-analysers

    • The Quanteon is a 4-laser flow cytometer that is configured with 25 independent silicon-based detectors (SiPM) to meet demanding sample panels. The SiPM detectors provide excellent sensitivity, stability, and a 7.2 log dynamic range! Very dim as well as very bright signals can be captured and gated in the same view, therefore eliminating the need for laborious trial-and-error PMT tuning procedures. Vesicles such as bacteria, platelets, or even extracellular vesicles as small as 0.1 μm can be detected (size based on polystyrene beads).
    • Excellent fluidics provides steady and consistent sample delivery, with a high reproducibility and exceptionally low CVs, even at high flow rates! The Quanteon provides volumetric fluidics absolute counts without the use of expensive beads and will, therefore, save time and money.
    • The Novosampler allows easy automated measurement with a tube rack (40 tubes) or various types of 96-well plates (however, no deep well), or even a 384-well plate. Additional cooling racks are available for the tube rack and plate loader.
    • The CFU has two identical Quanteons that can each serve as a backup in case of technical problems. Please note that there may be small differences in sensitivity and gain settings between the two systems!
    • Lasers (OBIS)
      • Violet: 404 nm, 100 mW
      • Blue: 488 nm, 100 mW
      • Yellow-green: 561 nm, 100 mW
      • Red: 640 nm, 100 mW

    • The BD FACSVerse cytometer is a compact research flow cytometer with three lasers (405, 488 and 640 nm) that has the ability to analyse up to 10 parameters (FSC, SSC, 8 colours (4 colours on the 488 laser, 2 colours on the 640 laser, and 2 colours on the 405 laser). Emission filters can be changed and the filters are automatically detected. Pulse processing (i.e. height, width, and area) can be used for all parameters. The vacuum-driven fluidics can make measurements at high (120 µl/min), medium (60 µl/min), and low (12 µl/min) flow rates.
    • In addition to manually delivering the sample to the BD FACSVerse cytometer for acquisition, the BD FACS Universal Loader can be used for multisampling using 30- and 40-tube racks (5 ml tubes) or multiple types of microtitre plates (96-well plates).
    • Samples are acquired (18-bits, 5 decades digitalization) using FACSuite software. Samples are saved as FCS3.0 files, allowing for compensation even after measurements have been made.
    • The cytometer performance is checked with the BD FACSuite CS&T research beads. Calibration with the CS&T beads allows automatic adjustments to take place, ensuring consistent values during each experiment on a daily basis. Reference or tube settings can be updated with the CS&T beads to correct instrumental variations.
    • For more specifications on lasers and detectors, please contact us. Our contact details can be found below.

    • The BD LSR-II cytometer is a research flow cytometer with 5 lasers that has the ability to analyse up to 20 parameters (FSC, SSC, and 18 colours). With these 5 lasers, we can measure different fluorochromes (2 on the blue laser, 3 on the red laser, 6 on the violet laser, 4 on the yellow-green laser and 3 on the UV laser). Bandpass and Dichroic filters can be changed with the help of one of the operators (this can be done only by the users themselves after having obtained permission from the facility). Pulse processing (i.e. height, width, and area) can be used for all parameters. The pressure-driven fluidics can make measurements at high (120 μl/min), medium (60 μl/min), and low (12 μl/min) flow rates (please note: fine adjustments can be made).
    • Samples are acquired (18 bits, 5 decades digitalization) using DIVA 8.0 software. Samples can be saved as FCS 3.0 or 3.1 files. Compensation afterwards is always possible.
    • For more specifications on lasers and detectors, please contact us. Our contact details can be found below.

    • The BD FACSymphony™ SORP is a conventional cell analyser. The CFU has a 30-parameter system with 8 lasers in total. To improve sensitivity and resolution, the Symphony is configured with 8 hot PMTs in the higher wavelength range.
    • The additional HTS (High Throughput System) allows automated measurements in 96-well plates.
    • Pulse processing (i.e. height, width, and area) can be used for all parameters. The pressure-driven fluidics can make measurements at high (120 μl/min), medium (60 μl/min), and low (12 μl/min) flow rates (please note: fine adjustments can be made).
    • Samples are acquired (18 bits, 5 decades digitalization) using DIVA 9.0 software. Samples can be saved as FCS 3.0 or 3.1 files. Compensation afterwards is always possible.
    • For more specifications on lasers and detectors, please contact us. Our contact details can be found below.

    • The Aurora is a spectral analyser that allows the detection of at least 35 different colours on the same cell type. A 40-parameter panel has recently been established (OMIP-069).
    • In a conventional flow cytometer, every fluorochrome is dedicated to just one channel, whereas the Aurora uses all 64 channels. There is no need to change the filters. In addition, this analyser provides a very high flexibility in choosing labels. As the whole spectrum is being analysed, unusual combinations, such as GFP and FITC, can be separated from each other.
    • The system is equipped with 5 flat-top lasers and has 64 channels for the detection of fluorescence. The electronics and APD detectors are very sensitive (low noise) and linear over a wide range (7 log decades).
    • A unique property is the ability to extract autofluorescence from cells, which allows the resolution of highly autofluorescent cells to be increased.
    • The small particle detection limit is approximately 110 nm for polystyrene beads (i.e. the same index as EVs) and 180 nm for silica beads.
    • Lasers:
      • UV: 350 nm, 20 mW
      • Violet: 405 nm, 100 mW
      • Blue: 488 nm, 50 mW
      • Yellow-green: 561 nm, 50 mW
      • Red: 640 nm, 80 mW

B-sorters

    • The MoFlo XDP is a high-speed sorter with 4 lasers (488 nm, 532 nm, 404 nm, and 642 nm lasers). It has the ability to analyse or sort up to 13 parameters (FSC, SSC, and 11 colours). According to the technical specification, it can achieve validated sort rates of up to 70,000 events/sec. In our experience, sorting rates of up to 25,000 events/sec are achieved, with good recovery, yield and viability when using the standard 70 nozzle. Although higher speeds (30,000-35,000 events/sec) are possible as well, the yield will be lower (because of the Poisson distribution). Four different populations can be sorted at the same time. Each population can be sorted using its own sort mode (Single, Purify, and Enrich).
    • In addition to the standard 70 nozzle, optional nozzle sizes (100 μm, 120 μm, and 150 μm) can be used to sort larger cells, which prevents clogging of the nozzle and creates better side streams. Although this will considerably reduce speed (no more than 8,000 cells/sec when using the 100 μm nozzle), the lower shear stress involved is gentler on the cells.
    • Multiple tube formats can be used for sorting when using the temperature-controlled SmartSampler. The sorted cells can be collected in multiple tube formats, such as Eppendorf tubes and 5 ml tubes. The Cyclone enables sorting in different plate formats (6, 12, 24, 48, 60 (Terasaki), 96, and 384 wells) and also on slides. Samples are acquired (32 bits – 5 decades) using Summit software. Samples are saved as FCS 3.0 files. Compensation afterwards is possible.
    • For more specifications on lasers and filters, please contact us. Our contact details can be found below.

    • The MoFlo Astrios is a high-speed sorter equipped with 7 lasers (355 nm, 405 nm, 488 nm, 532 nm, 561 nm, 592 nm, and 640 nm). The MoFlo Astrios has the ability to analyse or sort up to 32 parameters (FSC, 6 SSC, and 25 colours). According to the technical specification, it can achieve validated sort rates of up to 70,000 events/sec. In our experience, sorting rates of up to 25,000 events/sec can be achieved, with good recovery, yield, and viability when using the standard 70 nozzle. Although higher speeds (45,000 cells/sec) are possible as well, a lower yield will be achieved (because of the Poisson distribution). Six different populations can be sorted at the same time. Each population can be sorted using its own sort mode (Single, Purify, and Enrich).
    • In addition to the standard 70 nozzle, optional nozzle sizes (100 μm, 120 μm, and 150 μm) can be used to sort larger cells, which prevents clogging of the nozzle and creates better side streams. Although this will considerably reduce speed (no more than 8000 cells/sec when using the 100 μm nozzle),the lower shear stress involved is gentler on the cells.
    • Multiple tube formats can be used for sorting when using the temperature-controlled SmartSampler. The sorted cells can be collected in multiple tube formats, such as Eppendorf cups, and Eppendorf tubes of 5, 15 and 50 ml. The Cyclone enables (single cell) sorting in different plates (12, 24, 48, 60 (Terasaki), 96, 384 wells) and also on slides (customized format).
    • Samples are acquired (32 bits, 5 decades digitalization) using Summit software. Samples are saved as FCS3.0 files. Compensation afterwards is possible.
    • For more specifications on lasers and filters, please contact us. Our contact details can be found below.

    • The benchtop SH800S cell sorter allows sorting of a wide range of cell sizes for many applications, using the 70 μm, 100 μm, and 130 μm microfluidic sorting chips. This novel, chip-based design is fully integrated with comprehensive fluidics controls and advanced automation for set-up, acquisition, sorting, and analysis to make sorting less subjective, more precise, and easier to perform.
    • System software is intuitive and supports the sorting process into tubes, 2 ml Eppendorf tubes, and 96- and 384-well plates. The SH800S can sort 2 populations at the same time. The software generates FCS 3.0 and 3.1 files that can be exported to third-party analysis software.
    • The sorter is equipped with a Class II, Type A2 Biosafety Cabinet to provide the operator and the sorted cells with protection. The biosafety cabinet (CAS BioMAT 2) is custom designed for the SH800S.
    • The optical design offers four colinear excitation lasers (488 nm, 405 nm, 561 nm, and 638 nm) and six fluorescence channels. Although this allows the measurement of up to six different fluorochromes, the probe combinations have to be chosen carefully because the lasers cannot be separated in time. Therefore, some combinations, such as APC-Cy7 and PE-Cy7, are not possible.
    • For more specifications on lasers and detectors, please contact us. Our contact details can be found below.

Analysis software

  • For fast t-SNE clustering and many other applications

  • A very complete but also slightly complex program

  • Most user-friendly

  • Old-fashioned but still useful to some

  • The standard for DNA analysis

Contact

Geert Mesander
Geert Mesander Senior operator central flowcytometry unit