Equipment and Instruments
The FCF laboratory area is located on level 5, corridor 01 in room 01 and can be reached via the lift or staircase B1.
To access corridor 01, you need to have your service ID card activated. You will receive this after the instruction. If you have not yet been instructed, please use the bell.
FCF is usually open from Monday to Friday, 9 am until 5 pm.
You can find the User Regulations here.
Bench space
In addition to the devices for FACS analysis and cell sorting, users have access to some bench space where simple procedures such as adjusting cell densities or additional filtration can take place.
Among other things, a laminar flow hood, two centrifuges, pipettes, suitable plastic goods and standard buffers are available.
Equipment
The FCF allows access and use of the following equipment:
- ViCell XR Cell Viability Analyzer (Beckman Coulter)
- FACS-based Cell Analysis: Becton Dickinson (BD) FACS Canto II
- FACS-based Spectral Cell Analysis: Cytek Aurora
- FACS-based Cell Sorting: Becton Dickinson (BD) FACS Aria II
- FACS-based Cell Sorting: Becton Dickinson (BD) FACS Aria Fusion
- FACS-based Cell Analysis: Becton Dickinson (BD) LSR Fortessa (in special cases)
ViCell XR Cell Viability Analyzer (Beckman Coulter)
The FCF has a Cell Viability Analyzer with which absolute and live cell counts can be determined electronically
Camera-based determination of live cell count using Trypan Blue; cell viability indicated in percent, concentration and cell count
Cell types are defined by size, brightness, sharpness and spot brightness
Single measurement and 12-station autosampler
Specifications:
- Cell diameter = 3 - 70 μm
- Cell concentration = 5 x 104 - 1 x 107 Zellen/mL
- Sample volume = 0.5 - 2.0 mL
- Measuring time per sample = 2 minutes
Flow cytometry based cell analysis: Becton Dickinson (BD) FACS Canto II
BD FACS Canto II is maintained by the FCF and operated by the user.
The BD FACS Canto II analyser is equipped with three lasers (405nm, 488nm and 633 nm) with a configuration of 2+4+2 detectors.
This allows the detection of up to ten parameters:
- Two physical parameters, forward scatter and side scatter, give information about cell size and granularity/complexity.
- Eight PMTs allow the detection of fluorescence parameters.
The maximum sample acquisition rate is 10,000 events per second.
Access to Becton Dickinson (BD) LSR Fortessa equipped with four lasers (401nm, 488nm, 561nm, 640nm) is feasible if required to carry out the experiment.
Flow cytometry based cell analysis: Aurora (Cytek)
Aurora is maintained by the FCF and is open for all users with experience in FACS after proper instruction.
Aurora is a spectral analyzer equipped with five lasers (355nm, 405nm, 488nm, 561nm und 640 nm) and 64 detectors (16-16-14-10-8) as well as FSC and two SSCs.
Aurora allows the detection of more than 40 fluorescences simultaneously in one color panel and up to 62 parameters can be recorded:
- Three physical parameters (forward scatter and one side scatter each for the blue and the violet laser), give information about cell size and granularity/complexity.
- 64 optical detectors detect relative fluorescence intensities. Due to the high number of detectors, fluorescence in a wavelength range between 365nm and 829nm can be completely detected.
The maximum sample acquisition rate is 35,000 events per second.
In 2019, Aurora was acquired thanks financial support from the Clinic for Gastroenterology, Metabolic Diseases and Internal Intensive Care Medicine (Medical Clinic III) and the SFB TRR 57 and has been available to FCF users.
At the beginning of 2020, IZKF funds were invested to upgrade Aurora from a 3-laser system to a 5-laser system.
Flow cytometry based cell sorting: Becton Dickinson (BD) FACS Aria II
Please note: BD FACS Aria II is only operated by members of the FCF as a service to the user. The users have no access to this machine.
The FACS Aria machine consists of a FACS Aria II base. This base has been upgraded with Aria II components (FACS Aria II fluidics, sorting nozzles, sort chamber and optical cuvette).
It is equipped with three lasers (405nm, 488nm and 633nm) with a 2+5+2 detector configuration.
This allows the detection of up to eleven parameters:
- Two physical parameters, forward scatter and side scatter, give information about cell size and granularity/complexity.
- Nine PMTs allow the detection of fluorescence parameters.
The maximum sample acquisition rate is 25,000 events per second while sorting.
Simultaneous sorting of up to 4 distinct populations into tubes, or single cell sorting of a single population onto plates or slides is possible.
Water cooling or heating of the sample is available.
Flow cytometry based cell sorting: Becton Dickinson (BD) FACS Aria Fusion
Please note: BD FACS Aria Fusion is only operated by members of the FCF as a service to the user. The users have no access to this machine.
A BD FACS Aria Fusion is available for cell sorting. The facility's newest FACS device is integrated into a class 2 safety cabinet and is located separately in an S2 laboratory. It is now possible to sort S2 material, human samples or potentially infectious material in the FCF.
It is equipped with five lasers (375nm, 405nm, 488nm, 561nm und 640nm) with a 3+6+2+4+3 detector configuration.
This allows the detection of up to 20 parameters:
- Two physical parameters, forward scatter and side scatter, give information about cell size and granularity/complexity.
- 18 PMTs allow the detection of fluorescence parameters.
The maximum sample acquisition rate is up to 70,000 particles per second in analysis mode and it can sort at up to 25,000 particles per second.
Simultaneous sorting of up to 4 distinct populations into tubes, or single cell sorting of a single population onto plates or slides is possible.
Water cooling or heating of the sample is available.
Antibodies for compensation and isotype controls
- FCF has a small portfolio of antibodies, isotype controls, (fixable) viability markers and compensation beads. If required, the FCF will provide some aliquots of these for a small price.
- The available antibodies/reagents and pricing information can be obtained from the FCF.
Software Licenses
FCF has software licenses for the analysis of FACS data, which are lent on a daily basis. The available and pricing information can be obtained from the FCF.
- FlowJoTM v10
- FCS Express 7, DeNovo Software