Core Unit FACS

Head:
Prof. Charlotte Esser tl_files/bilder/mail.gif
Phone: +49 (0)211-3389-253

Academic staff:
Dr. Nadine Teichweyde (Training, Advice, Assistance) tl_files/bilder/mail.gif
Phone: +49 (0)211-3389-252

Profile

Cell sorting and analyses are used for many research questions of the IUF, including

  • characterization of cells by surface and intracellular staining
  • Calcium Influx
  • microbiome profiling
  • cell cycle analyses, cell proliferation and cytotoxicity analysis
  • detection of reactive oxygen species (ROS)


The core unit serves these needs with their FACS Canto II and a FACS Aria III instruments from Becton-Dickinson
The FACS Canto II, a flow cytometer for analysis, is equipped with three lasers (405 nm, 488 nm, 633 nm) and eight fluorescence channels for multi-parameter analyses.

The FACS Aria III is a high-speed cell sorter, equipped with four lasers (405 nm, 488 nm, 561 nm, 633 nm) and sixteen fluorescence channels. The FACS Aria III is able to perform 4-way sorts and single cell sorting directly into 6, 12, 24, 48,96 well plates with the possibility of Index Sorting, which allows a retrospective identification of each single cell’s phenotype.

As software, we use FlowJo™ and FACSdiva™. Together, the FACS Unit allows addressing a wide range of analytical questions and sorting approaches. Moreover, the Core Unit is an S2 laboratory and Biosafety-level 2 laboratory

Dr. Teichweyde, who also performs cell sorts on the FACS Aria III on request, offers training, help and assistance for users. First time users should contact her with details of the experimental plans, including S1, S2 or biosafety requirements.

Recommended Literature:

Cossarizza…Esser… et al.  Guidelines for the use of flow cytometry and cell sorting in immunological studies. Eur J Immunol. (2017) 47:1584-1797.

Jamie A. Lee et al. MIFlowCyt: The Minimum Information About a Flow Cytometry Experiment. Cytometry Part A. (2008) 73:926-30.

Esser C, Hochrath K, Teichweyde N, Krutmann J, Chang HD. Beyond sequencing: fast and easy microbiome profiling by flow cytometry. Arch Toxicol. (2019) 93:2703-2704.

https://currentprotocols.onlinelibrary.wiley.com/

More information on fluorescent colors and their spectra:

https://www.bdbiosciences.com/en-us/applications/research-applications/multicolor-flow-cytometry/product-selection-tools/spectrum-viewer

Fluorochrome/Laser Reference Poster:

https://www.bdbiosciences.com/documents/multicolor_fluorochrome_laser_chart.pdf