The central Flow Cytometry Core Facility (FCF) provides all scientists across all institutes with access to established and innovative methods and instruments for their respective scientific questions. The strengths of flow cytometry lie in the rapid examination of a wide range of parameters of cells or other particles in suspension. This enables detailed analysis at the single-cell level and thus also the performance of complex experiments and the use of heterogeneous samples with good statistical validation of the results. We offer flow cytometric data acquisition/analysis of multicolor samples on our analyzers as well as sorting of up to four target populations simultaneously from one sample on the sorters. Of note, the entire Core Facility is approved according to current S2 regulations by the Regierungspräsidium Tübingen.
About us
Services
Consulting
Advice for panel design and analysis
Cell Sorting
Sort consultation
Basic cell sorts
Complex sorts under sterile conditions
Sorts into 96- or 384-well plates
Cell sorting of infectious samples
Flow Cytometry Analysis
Flow cytometry analyses with up to 16 colors
Spectral analyzer measurements and analyses
ImageStream measurements
Training
Training for users
all analyzers & ImageStream
Software
Training and use in
FlowJo, DIVA, IDEAS, support with Cytolution
Team
Dr. Anna Stanger
Head of FCF Core Facility
Dr. Kristin Bieber
Technial Head
Pardis Khosravani
Scientist
Sabrina Grimm
Technician
Salina Amos
Technician
Equipment
Cell Sorter
Cell sorting is performed by FCF staff on a BD FACS Aria cell sorter equipped with four lasers (355 or 402, 488, 561, 640 nm excitation wavelengths) and 14 fluorescence channel detection. The cell sorter is able to perform high-speed (up to 20,000 events/sec), high-purity (up to 99.9%) sterile sorts (biosafety level 2) into different types of devices (tubes, plates or slides). The cell sorter is operated by FCF staff during the extended business hours. We perform basic cell sorts (e.g. lymphocytes, DCs, cells lines, transfected cells,..), but also complex sorts under sterile conditions (e.g. use of near UV-laser for stem cell side population of tumor tissue), sorts into 96- or 384-well plates with defined cell numbers (even single cells) and more.
Images are courtesy of BD Biosciences and Sony.
BD FACS Aria IIIu
- 4 laser (402 nm, 488 nm, 561 nm, 640 nm)
- 14 fluorescence channel detection
- 4-way sorting
- Bulk- and single cell sorting
- Index sort
- Nozzle size: 70 µm, 85 µm, 100 µm
SONY MA900 Multi-Application
- 4 laser (405 nm, 488 nm, 561 nm, 640 nm)
- 12 fluorescence channel detection
- 4-way sorting
- Bulk- and single cell sorting
- Index sort
- Micro fluidic chip-based design: 70 µm, 100 µm, 130 µm disposable sorting chips
DFG-funded (project number: 424726817)
Analyzer
The FCF is equipped with four advanced benchtop analyzers, which have up to four lasers (488, 561, 640 and 405 nm excitation wavelengths) and 16 fluorescence channels. Researchers can be trained by the FCF staff in order to use the machines independently without an operator. We also provide detailed advice and support on the choice of fluorophores, equipment and protocols.
Images are courtesy of BD Biosciences.
BD LSR Fortessa
- 4 laser (405 nm, 488 nm, 561 nm, 640 nm)
- 18 color analyzer
BD LSR Fortessa + HTS Loader
- 4 laser (405 nm, 488 nm, 561 nm, 640 nm)
- 18 color analyzer
- Autosampler
DFG-funded (project number: 424726769)
BD FACS Canto II
- 3 laser (405 nm, 488 nm, 640 nm)
- 8 color analyzer
BD FACS Lyric
- 4 laser (405 nm, 488 nm, 561 nm, 640 nm)
- 10 color analyzer
- Autosampler
Spectral Analyzer
The CYTEK™ Aurora allows separation of fluorochromes with very close emission profiles. This enables the use of a wide range of new fluorochrome combinations. The machine is equipped with UV, violet, blue, yellow/green and red lasers and provides up to 64 fluorescence channels. A plate sample loader is part of the assembly. The AURORA is available for independent use following a training by the FCF staff.
Image courtesy of CYTEK™.
CYTEK™ AURORA
- 5 laser (355 nm, 405 nm, 488 nm, 561 nm, 640 nm)
- Up to 64 fluorescence channels
- Autosampler
Reference controls operation protocol
DFG-funded (project number: 424726600)
Imaging Flow Cytometry
Imaging flow cytometry combines the features of flow cytometry and fluorescence microscopy and overcomes the limitations of both techniques. The CYTEK™ Amnis™ ImageStream™ Mark II is equipped with violet, blue, yellow/green and red lasers, twelve image channels, two highly sensitive CCD cameras, three objective lenses and a tool for extended depths of field (EDF). The CYTEK™ Amnis™ ImageStream™ Mark II is operated by the FCF staff. A preliminary consulting is mandatory to clarify the overall goal of the experiment or project. We offer advice and support for sample preparation and are happy to perform the data analysis together with the researchers also possibly including data analysis and figure preparation for publications.
Image courtesy of CYTEK™.
CYTEK™ Amnis™ ImageStream™ Mark II
- 4 laser (405 nm, 488 nm, 561 nm, 642 nm)
- 12 image channels
- 2 CCD camera systems
- MultiMag (20x, 40x, 60x)
- Extended depths of field (EDF)
FAQ
How do I get started?
For using our services, you will need a user account on our booking system PPMS.
Please create your user account here.
For cell sorting requests:
No training required, since FCF staff handles the devices.
Please send us an email with the following information:For using our analyzers:
Basic training is mandatory! Even if you have already worked with a flow cytometer before!
Please send us an email with the following information:- Do you bring experience in flow cytometry? On which instrument/s? How many colors?
- Do you request for training only or will you measure your samples at our facility after completing the training?
- User registration sheet BD FACS analyzer
How do I book/cancel sessions?
Booking
In order to book an appointment, you need to have a user account on our online booking system PPMS.
For sort appointments: Since cell sortings are conducted by FCF staff only, please book a consultation first. We discuss new cell sorting experiments with you every Wednesday from 10:00-12:30.
FCF-approved users can simply go to the online calendar and book their preferred time slot.
Cancelation
A booked session can be canceled free of charge up to 24 hours before the actual start of the session.
Thereafter, the usage fees will apply to our late cancelation policy. If a user does not show up through the entire booked session, usage fees will be calculated according to booked time.
How do I prepare my samples for a sort?
Please bring an unlabeled sample plus single staining controls to properly set up the experiment.
It is essential to filter the cells through a 40 µm cell strainer prior measurement.
For more detailed information follow our detailed guidelines.
Which controls do I need?
Beside an unlabeled sample and single staining controls, we recommend to work with FMOs (fluorescence minus one) in order to set up the experiment properly.
How do I prepare my compensation/reference controls?
In principle, you can either use beads or cells.
Usage of beads is recommended if cell number is limiting, the antigen is dimly expressed or the positive population is rare.
Recommendations:
- For CYTEK™ Aurora: Treat your reference control samples in the exact same way as your experimental samples in order to get a clean and accurate signature of the fluorophore.
- For Amnis ImageStreamx Mark II: Use cells, not beads, as reference controls for a more accurate calculation of the compensation matrix.
How much will it cost?
Please contact fcf-berg@med.uni-tuebingen.de for the full price list.
Why can't I see all the other analyzers in my PPMS account? I have completed my basic training.
You are only able to book sessions on instruments you have been trained on. Every single instrument in our facility requires training. Please send us an email if you want to use another instrument than the one you were introduced to in your basic training.