Cytometry and Cell Sorting Core Unit
Aim of the Cytometry and Cell Sorting Core Unit is to provide interested scientists access to latest equipment of flow cytometry.
- 4 Analyser (LSR Fortessa-A3, Quanteon, LSR Fortessa, CantoII till the end of 2024, Aurora at the end of 2024)
- 2 Sorter (AriaIIIu und AriaFusion)
- 1 Imaging Analyzer (ImageStream)
The use of the analyzers is possible under your own responsibility after prior training and acceptance of the terms of use (also offered as a service). The operation of the FACS AriaIIIu and the FACS AriaFusion will be done by the employees of the facility.
Services
- Cell Sorting with FACS AriaIIIu and FACS AriaFusion
- Cell Analysis with FACS CantoII and LSR Fortessa
- Methodological and technical support
- Support for the establishment of new applications
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- Multicolor-Analysis
- Surface markers
- Intracellular markers
- Fluorescence proteins
- Functional and Immuno-Assays
- Apoptosis
- Cytokine detection
- Cell cycle analysis
- Side population
- Multicolor-Analysis
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- Training on FACS CantoII and LSR Fortessa and, for people with longterm FACS-experience on FACS AriaIIIu and FACS AriaFusion
Cell Analysis
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FACS Canto II
By FACS Canto II FACS measurements can be carried out independently or by the core facility staff. The device is equipped with the following lasers:
- 407 nm: violet laser
- 488nm: Blue Laser
- 633 nm: Red Laser
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FACS LSR Fortessa
The LSR Fortessa expanded as a new flow cytometer FACS Sorting Core Unit. The device allows, in addition to the blue, violet and red laser a yellow-green laser for better detection with the use of PE-dyes and red fluorescent proteins. Up to 17 colorants are used in a sample.
By FACS LSR Fortessa FACS measurements can be carried out independently or by the core facility staff. The device is equipped with the following lasers:
- 405 nm: violet laser
- 488nm: Blue Laser
- 561 nm: green laser
- 640 nm: Red Laser
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Cell Sorting
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FACS Sorting - The principle
1. Cells are harvested by a sheath flow as beads strung on a string (hydrodynamic focusing)
2. In the laser intersection the analysis of the cells takes place.
3. The nozzle separates the jet into individual drops and charges the drops to be sorted on.
4. The charged drops are deflected in an electric field, and fall into the appropriate receptacle.
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Sorting: duration
The following tables show how long a sorting operation. These are minimum hours, 20 minutes should be allocated for the setting of the device. The duration varies with the size of the cells.
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Sorting: multicolor experiments
Examples for multicolor analysis
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Sorting: different sample collections
The following options are possible with the FACS Aria III and FACS Aria Fusion:
1) separated into 2 populations in 15 ml tubes
2) seperated into up to 4 populations in FACS-tubes or small reaction tubes
3) filing in microtiter plates (for example, 96-well)
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FACS AriaIllu
This Cytometry one or more cell populations from the suspension can be sorted out (Note: not all cells can be sorted well, sometimes the populations are very sensitive). The sorted cells can be cultivated further.
The cell sorting on FACS AriaIIIu is handled by employees of the Core Unit. The device is equipped with the following lasers:
- 407 nm: violet laser
- 488nm: Blue Laser
- 561 nm: green laser
- 633 nm: Red Laser
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FACS AriaFusion
The device is equipped with the following lasers:355 nm UV laser
- 405 nm: violet laser
- 488nm: Blue Laser
- 561 nm: green laser
- 640 nm: Red Laser
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Further Equipment
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Imaging Flow Cytometer
Technical data:
Laser:
- 488 nm blue laser (high-power)
- 642 nm red laser
- 785 nm far red laser
- 375 nm UV laser
- 405 nm violet laser
- 561 nm green laserSpeed:
- up to 200 events per secObjective
-20x, 40x und 60x magnicficationSample characteristics:
- Volume: 20-200yl
- Utilization efficiency: up to 95%
Registration
The FACS Sorting Core Unit is open to all researchers at the UKE and other interested researchers. Booking is done through the provided Internet booking system. New users call directly to the specified contact person.
There is usage fee per hour:
• FACS Canto II: 17 € per hour
• FACS LSR Fortessa € 22 per hour
• FACS AriaIII 60 € hour
• FACS AriaFusion 60 € hour
Contact
Location:
N27 (Campus Forschung)
4. Etage Raum 61.1
- Core manager
- Cytometry and Cell Sorting Core Unit
- Deputy Head
- Cytometry and Cell Sorting Core Unit
- Chemical-technical assistant
- Cytometry and Cell Sorting Core Unit