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About
We have extensive experience with flow cytometry in basic and translational research, ranging from single cell phenotyping and sorting to extensive immuno-profiling in clinical trials. We help in-house and external researchers to obtain high quality flow cytometry results through advice, support, state-of-the-art equipment and (computational) data-analyses tools.
Expertises and services
Let us enhance your research
Researchers, but also knowledge institutes and companies, approach us for our expertise and equipment. For example for:
- access to equipment
- advice on panel design
- advice on data analysis
- training in flow cytometry
Let us enhance your research
Researchers from Radboudumc and beyond, but also knowledge institutes and companies, approach us for our expertise and high-end equipment.
Examples of how flow cytometry can be used:
- (Immune) cell phenotyping
- Transfection efficiency
- Cell counting
- Cell sorting based on fluorescent proteins
- Multi-well single cell sorting in 6-384 well plates for single analysis applications
- Nucleic acid analysis/Cell cycle analysis
- FRET measurements
- Small particles analysis and sorting
- Cell function analysis (calcium efflux)
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"At the RTC there is always someone available with knowledge of flow cytometry, willing to help with possible questions" (Mercurna - translated from Dutch)
“The RTC flow cytometry of our hospital has significantly facilitated my research by providing high-quality training, state-of the art flow cytometry facilities and trouble-shooting from knowledgeable technicians and researchers. Everyone is willing to help and assist in all phases of research including experimental design, execution (cell sorting) and data analysis.” (Rheumatology)
“I use the RTC flow cytometry very often, and more recently almost on a weekly basis. It started with the technical need for user-friendly flow cytometers and now I'm even reaching out for help with panel building and decision making. On top of this, I also make use of the sorting assistance for sample preparation in complex experiments. I can say I'm a frequent and satisfied user.” (Cell biology)
What is flow cytometry?
Flow cytometry allows the interrogation of few to millions of single cells or particles.
read moreWhat is flow cytometry?
Flow cytometry is a sophisticated technique that allows interrogation of few to millions of single cells or particles.
Multiple physical characteristics of each individual cell/particle are simultaneously measured, and expression levels of antigens can be detected using fluorescently labelled antibodies. Measured characteristics include amongst others the relative size, granularity/internal complexity, and relative fluorescence intensity of the cell/particle.
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A flow cytometer is composed of three main systems: fluidics, optics and electronics. The fluidics transports the particles in a stream to the laser beam for interrogation. The optics consists of lasers to illuminate the particles in the sample stream and optical filters to direct the resulting light signals to the appropriate detectors. The electronics convert the detected light signals into digital signals that can be processed by the computer.
Monoclonal antibodies tagged with a fluorescent dye can be used to identify a particular cell type based on binding to individual surface, cytoplasmic, or nuclear antigens. Different fluorochromes can be used simultaneously to distinguish sub-populations. Dyes, which bind directly to cell components such as DNA and RNA, are used as well.
The flow cytometry technology provides a powerful and sensitive tool for investigating the structure and function of cells, bacteria and other particles. These measurements can be made at flow rates of 100-20,000 cells per second.
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Cell sorting allows us to capture and collect cells of interest for further biochemical, or functional analyses. Cells can selectively be removed from a mixed population based on specific physical characteristics, for example, size or fluorescence. A voltage charge is applied to drops containing the cells that meet the predefined sorting criteria. Positively and negatively charged plates deflect the charged drops into collection devices, such as tubes or microscope slides, depending on the charge polarity. As sorting is performed under sterile conditions, sorted cell fractions can be cultured after sorting.
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Proper interpretation of flow cytometry data is a key factor and requires some experience and insight. Using available graphical presentation tools like dot plots and density graphs are common in the field of flow cytometry. New tools like dimension reduction and cluster analysis give more insight in the relation between markers and cell populations and are powerful tools for high dimension flow cytometry data. There are a wide variety of sophisticated computer programs to assist in the display and analysis of flow data.
Using our equipment and mandatory steps
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Read more about training and booking options before making use of the Booking platform.
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Instructions
It is recommended to get familiar with our equipment and flow cytometry before requesting an instruction on the equipment.
Users who have been trained on our flow cytometry equipment are allowed to use the equipment without assistance. Individual and group training in the use of our equipment is organized upon request.
- Training on the analyzers is provided once a month and can be attended upon request.
- For cell sorters it is optional to be trained yourself, but operators at our technology center are always willing to do the sorting for you. Training is dependent on the amount of sorts you need and the frequency of your experiments.
After training you are allowed to book our equipment via the booking website. Analyzers can be booked at any open time-slot, as you operate them yourself. Assistance can generally be provided from Mondays to Thursdays at working hours. Sorters can be booked during working hours and will be operated by our staff or by yourself after training.
We are located in the F-building on the third floor, route 818.
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Visit the Radboud's Booking platform, section "Flow Cytometers RTC".
visit website
Analyzers
Our conventional flow cytometry analyzers and spectral analyzer allow for simple or more advanced cell analysis. The analyzers are situated in an MLI biohazard room.
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This advanced flow cytometer presents optimal excitation and emission, minimizing light loss and maximizing sensitivity.
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CytoFLEX LX (21 colors)
This flow cytometer is an advanced system presenting optimal excitation and emission, minimizing light loss and maximizing sensitivity. The fully activated instrument includes three fluorescent channels from the 488 nm (Blue) laser, three from the 638 nm (Red) laser, five from the 405 nm (Violet) laser, five from the 561 nm (Yellow Green) laser, three from the 355 nm (UV) laser and two from the 808 nm (Infrared) laser.
Acquisition is possible with tubes and 96 (deep) well plates. Volumetric sample injection allows absolute cell counting. The system operates with Cytexpert software. Generated FCS files can be further analyzed with Kaluza or Flowjo.
Book this analyzer and/or download laser and filter configuration through Radboud's Booking platform, section "CytoFLEX LX".
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This flow cytometer is as sensitive and reliable as its big brother, the CytoFLEX LX, but useful for less complex panels.
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CytoFLEX (13 colors)
This flow cytometer includes five fluorescent channels from the 488 nm (Blue) laser, three from the 638 nm (Red) laser and five from the 405 nm (Violet) laser.
Acquisition is possible with tubes and 96 (deep) well plates. Volumetric sample injection allows absolute cell counting. The system operates with Cytexpert software.
Generated FCS files can be further analyzed with Kaluza or Flowjo.Book this analyzer and/or download laser and filter configuration through Radboud's Booking platform, section "CytoFLEX" (for more complex panels, we recommend the CytoFLEX LX).
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This flow cytometer combines flowcytometry data with image parameters and is especially equipped for small particle measurement like EVs.
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Cellstream
This flow cytometer combines flow cytometry data with image parameters for better data clean up.
The machine is especially equipped for small particle measurement like EVs.Book this analyzer and/or download laser and filter configuration through Radboud's Booking platform, section "Cellstream".
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This flow cytometer can be used for analysis of advanced multi-color panels up to 50 colors, refer to the block "Cell sorters" for further specifics.
Cell sorters
Our Aurora cell sorter can measure up to 40 colors and sort 6 populations. The CytoFLEX SRT sorter is placed in an MLII lab allowing for sorting of virally transduced cells.
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This cell sorter is equipped with 5 lasers. It is able to use previously undistinguishable (of similar) colors and make panels up to 50 colors.
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This cell sorter has 4 lasers and can be used mainly for GMO material, both MLI and MLII.
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CytoFLEX SRT
This cell sorter can be used mainly for GMO material, both MLI and MLII.
This device has 4 lasers (405, 488, 562 and 633) and 15 fluorescent channels. 4 way sorting and 96 or 364 wells plates are possible, nozzle size is 100µm
Users are allowed to operate the machine themselves after an introduction.
The following fluorescent proteins are suitable for sorting with the SRT:
- GFP; YFP; CFP; RFP; mCherry; dsRed and many other
These dyes are usually used in combination with a viability dye suitable for one of the lasers.
Book this analyzer and/or download laser and filter configuration through Radboud's Booking platform, section "CytoFLEX SRT".
Software
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We offer Kaluza analysis software (Beckman Coulter) as a flow cytometry analysis tool in the Radboudumc network. This software can process files generated on CytoFLEX and AuroraCS. This software is easy to use and will accommodate many research questions.
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Flowjo (BD) can be used on the data analysis computers in our RTC. This software can process files generated on CytoFLEX and AuroraCS. This software is more advanced and requires a bit more training.
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Omiq is a program for more advanced data analysis, this can be used upon request. Pre-analysis and compensation are normally performed before transferring to Omiq. Omiq is strong in cluster analysis and dimension reduction for large panels.
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Cytek Cellstream can be used on the data analysis computers in our technology center. This software is specifically for cellstream data.
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Cytek Spectroflo can be used on the data analysis computers in our technology center. This software processes raw and unmixed datafiles from AuroraCS.
Also see...
GMO information
We comply with the GMO (genetically modified organisms) rules and regulations of Radboudumc. GMOs for analysis and sorting should be mentioned in the central GMO license. MLI cells can be measured and sorted in our technology center. MLII cells for analysis should be fixed with 1% PFA. MLII cells for sorting are registered in our local logbook. Please contact our technology center for more information.
Events
Do you want to increase your knowledge of flow cytometry? Every year, the Radboudumc organizes several events in which flow cytometry is central, such as the 'Meet the expert' sessions or symposia on, for example, (high-dimensional) data analysis. When available, these events appear in the general research agenda.
see pageUseful links
- Fluorofinder - advanced spectraviewer, panel builder and dye information. Flow cytometer configurations are available in the database.
- OMIPS - advanced published predesigned panels.
- BioLegend flow cytometry tools - info on fluorophores and other tools.
- ISAC - provides a key journal on flow cytometry issues, optimizations and panels.
Organization and people
Getting here
Directions
Connected to
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Thessa Scheele research assistant (e.g. technician, data analist) analist Laboratoriumgeneeskunde
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Rob Woestenenk hoofdanalist
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Tjitske Duiveman-de Boer applied translational researcher tumor immunologie
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Diede van Ens laboratoriumspecialist
