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.
Publications Flow Cytometry
Martins S, Hacheney I, Teichweyde N, Hildebrandt B, Krutmann J, Rossi A: Generation of an induced pluripotent stem cell line (IUFi001) from a Cockayne syndrome patient carrying a mutation in the ERCC6 gene. Stem Cell Res 55: 102456, 2021. doi: 10.1016/j.scr.2021.102456
Tigges J, Bielec K, Brockerhoff G, Hildebrandt B, Hübenthal U, Kapr J, Koch K, Teichweyde N, Wieczorek D, Rossi A, Fritsche E: Academic application of Good Cell Culture Practice for induced pluripotent stem cells. ALTEX 2021. doi: 10.14573/altex.2101221
Merches K, Schiavi A, Weighardt H, Steinwachs S, Teichweyde N, Förster I, Hochrath K, Schumak B, Ventura N, Petzsch P, Köhrer K, Esser C: AHR signaling dampens inflammatory signature in neonatal skin γδ T cells. Int J Mol Sci 21(6): 2249, 2020. doi: 10.3390/ijms21062249
Häselbarth L, Ouwens DM, Teichweyde N, Hochrath K, Merches K, Esser C: The small chain fatty acid butyrate antagonizes the TCR-stimulation-induced metabolic shift in murine epidermal gamma delta T cells. EXCLI J 19: 334-350, 2020. doi: 10.17179/excli2020-1123
Esser C, Hochrath K, Teichweyde N, Krutmann J, Chang H-D: Beyond sequencing: fast and easy microbiome profiling by flow cytometry. Arch Toxicol 93(9): 2703-2704, 2019. doi: 10.1007/s00204-019-02527-1
Cossarizza A, … Esser C, …. et al.: Guidelines for the use of flow cytometry and cell sorting in immunological studies. Eur J Immunol 47: 1584-1797, 2017. doi: 10.1002/eji.201646632
Jamie A. Lee et al.: MIFlowCyt: The minimum information about a flow cytometry experiment. Cytometry Part A 73: 926-930, 2008. doi: 10.1002/cyto.a.20623
More information on fluorescent colors and their spectra:
Fluorochrome/Laser Reference Poster: