Instrumentation
The Core Facility is equipped with two cell sorters, one research cell analyzer and a confocal laser scanning microscope.
Cell Sorters
BD FACSAriaIII Cell Sorter (Room 067.000 20 014.02-1)
The BD FACSAriaIII Cell Sorter has an optically gel-coupled cuvette flow cell. The true fixed laser alignment system minimizes startup time, improves experiment-to-experiment reproducibility, and enables automated daily quality control.
The FACSAriaIII can be operated with a 70, 85 or 100 µm nozzle depending on the sample type. Nozzles are readily accessible and easy to change.
The Cell Sorter is equipped with 5 lasers:
| Laser | Wavelength (nm) | Output power (mW) |
| 1 | 488 | 20 |
| 2 | 561 | 50 |
| 3 | 633 | 18 |
| 4 | 405 | 50 |
| 5 | near UV 375 | 7 |
The collection optics of the BD FACSAriaIII is set up to measure 18 color parameters + FSC/SSC. The standard configuration with band-pass and long-pass filters can be found in PPMS in form of a table under My Documents. Numerous swappable filters are available and can easily be installed.
The FACSAriaIII is able to sort 4 subpopulations at a time. Cells for sorting can be delivered in 12 x 75 mm, and 15 ml tubes. Cells can be sorted into 15 ml tubes (2-way sorting) or into 1.5 ml and 12 x 75 mm tubes (4-way sorting). The Automatic Cell Deposition Unit (ACDU) enables sorting onto slides and into plates (6, 24, 48, 96, and 384-well plates). Sample input and collections tubes, plates, and slides can be temperature controlled to 4, 20, 37 and 42°C respectively.
The BD FACSAriaIII is exclusively operated by the CAnaCore staff. The intended use of the device is cell sorting! Registered users have to fill out a booking form in PPMS before they are able to make a sorting appointment on the FACSAriaIII. A "Cell Sorting Guide" which can be found in PPMS under My Documents provides the basic information on what to consider when setting up a sorting experiment.
Beckman Coulter MoFlo Legacy Jet-in-Air Cell Sorter (Room 067.000 20 014.02-1)
Beckman Coulter (formerly Dako) MoFlo Legacy is a Jet-in-Air High Speed Cell Sorter. The modular platform offers flexibility regarding the configuration of the optical bench.
MoFlo is routinely operated with a 100 µm nozzle at 20 psi. Also available is a 70 µm nozzle for "high speed sorting" and a 150 µm nozzle for large cells or particles.
The Cell Sorter is equipped with 5 lasers:
| Laser | Wavelength (nm) | Output power (mW) |
| 1 | 488 | 10 - 200 |
| 2 | 561 | 10 - 100 |
| 3 | 405 co-linear with 640 | 10 - 100 |
| 4 | 640 | 10 - 100 |
Theoretically MoFlo is capable to detect 10 color parameters + FSC/SSC. Since the compensation matrix in Summit, however, is able to handle only 8 color parameters the panel design should not exceed more than 8 antibodies/fluorochromes. The standard filter configuration with band-pass and dichroic mirrows of MoFlo can be found in PPMS under My Documents. Numerous swappable filters are available and can easily be installed.
MoFlo is equipped with a 4-way sorting module which facilitates the sorting of up to 4 subpopulations. Cells for sorting can be delivered in 1.5, 12 x 75 mm, 15 ml, and 50 ml tubes. Cells can be sorted into almost all saleable tubes (1.5 ml, 12 x 75 mm, 15 ml, and 50 ml tubes for 2-way sorting; 1.5 ml, 12 x 75 mm tubes for 4-way sorting). The robotic arm “Cyclone” enables sorting into multi-well plates (6-well to 384-well), in some cell culture vessels (Petri dishes 5 – 10 cm in diameter), chamber slides, and onto microscope slides.
MoFlo Legacy is exclusively operated by the CAnaCore staff. The intended use of the device is cell sorting. Registered users have to fill out a booking form in PPMS before they are able to make a sorting appointment on MoFlo. A "Cell Sorting Guide" which can be found in PPMS under My Documents provides the basic information on what to consider when setting up a sorting experiment.
Research Cell Analyzer
BD LSRII Analyzer (Room 067.000 20 014.01)
The BD LSRII benchtop analyzer is designed for advanced analytical research applications using digital signal processing and a flow cell with gel-coupled cuvette. Data acquisition is performed by using the BD FACSDiva Software. The BD LSRII is a self-operated system. Potential users must undergo a hands-on training before they are able to book and operate the benchtop analyser. Trainings will be given on inquiry by using the corresponding request form in PPMS.
The LSRII is equipped with 4 lasers:
| Laser | Wavelength (nm) | Output power (mW) |
| 1 | 488 | 20 |
| 2 | 633 | 18 |
| 3 | 561 | 100 |
| 4 | 405 | 25 |
The collection optics of the LSRII is set up to measure 15 color parameters + FSC/SSC. The standard configuration with band-pass and long-pass filters of the BD LSRII can be found in PPMS under My Documents.
Cells for analyzing must be prepared in 12 x 75 mm tubes suitable for Flow Cytometry to fit onto the sample injection port.
All users must stand to the BD LSRII user guide line which can be found in PPMS under My Documents! In addition all users should be aware of the Startup and Shutdown procedure of the LSRII.
Confocal Laser Scanning Microscope
Olympus FV1000 Confocal Microscopy System (Room 067.000 20 014.03)
The Olympus FV1000 IX81 is a laser scanning confocal microscope built for life cell imaging. The imaging platform is operated using Fluo-View-FV1000 2.0b.
The FV1000 is equipped with an environmental chamber which allows the user to maintain stable experimental conditions like 37°C, 90% humidity and 5% CO2 concentration for long-term life imaging if needed.
The FV1000 IX81 is equipped with 4 lasers:
| Laser | Wavelength (nm) | Output power |
| 1 | 458, 488, 515 | adjustable |
| 2 | 635 | adjustable |
| 3 | 559 | adjustable |
| 4 | 405 | adjustable |
All users are required to undergo a intensive training before using the microscope. Trainings are giving on inquiry. A corresponding request form can be found in PPMS.
FlowJo/SeqGeq Work Stations
CAnaCore has two powerful work stations for data analysis using either FlowJo or SeqGeq. Those work stations are bookable via PPMS and are of no charge.