Flow cytometry

Various flow cytometers and sorters are available at the Fraunhofer IZI. We will be happy to help you select the equipment for your experiments.

For examples, the following devices are available.

• Specifications: Laser: 405 nm, 488 nm, 633 nm, 8 fluorescence channels
• Assays: measurement of surface markers, cell differentiation, live-dead assays, human & murine samples, primary cells/cell culture (e.g. PBMCs, CAR-T, CAR-NK, various cell lines), primary cells (primarily mouse, partially human, rarely rat), blood cells or single cell suspensions of various organs, immune status, intracellular measurements (cytokines, transcription factors, proliferation, etc.), cell culture samples
• Reference projects
  • Allometric Dose Retranslation Unveiled Substantial Immunological Side Effects of Granulocyte Colony–Stimulating Factor After Stroke
  • Flow cytometric characterization of brain dendritic cell subsets after murine stroke
  • High-dosage granulocyte colony stimulating factor treatment alters monocyte trafficking to the brain after experimental stroke
  • Isolation and Flow Cytometric Analysis of Immune Cells from the Ischemic Mouse Brain

• Specifications: 6-way sorter, lasers: 488, 561, 640, 2ps pulse laser (375, 488), 8 fluorescence channels (expandable) / octagon system (BD)
• Assays: Live-dead staining, apoptosis/necrosis, mitochondrial activity, surface markers for cell identity and characterization, surface markers + intracellular for determination of activation (immune cells, pro/anti-inflammatory), detection of transcription factors as proof of identity and function, and differentiation status (pluripotency, lung, etc), detection of transgene expression in transfections and transductions (mRNA, plasmids, viral vectors), sorting of cells via flow cytometry

• Specifications: 3 lasers (405 nm, 488 nm, 638 nm), 13 fluorescence channels (450/45, 525/40 (2), 585/42, 610/20 (2), 660/10 (2), 690/50, 712/25, 780/60 (3)), high-throughput measurements possible
• Assays: internalization; 10-color immunophenotyping; senescence assays; cell cycle analysis; intracellular cytokine staining
• Reference project
  • Transcriptomic signatures reveal a shift towards an anti-inflammatory gene expression profile but also the induction of type I and type II interferon signaling networks through aryl hydrocarbon receptor activation in murine macrophages

Specifications: 3 lasers (405 nm, 488 nm, 638 nm), acquisition of high-resolution images of each cell directly in the flow (bright and dark field (SSC), as well as up to 12 fluorescent markers), high-throughput measurements possible

Assays: Presentation on Amnis® ImageStream®X Mk II

• Specifications: 2 lasers (488 nm, 640 nm), 4 fluorescence channels simultaneously (existing filters: 533/30, 585/40, 670LP, 675/25), high-throughput measurements possible
• Assays: measurement of beads, cells, bacteria, apoptosis assay, binding curves (EC50 value calculation), transfection efficiencies
• Reference project
  • Pinpointed Stimulation of EphA2 Receptors via DNA-Templated Oligovalence

• Specifications: 3 lasers (405 nm, 488 nm, 640 nm), 8 fluorescence channels, high-throughput compatibility through Buffer Supply Station, automatic staining, measurement in express and user-defined mode possible
• Assays: Cell count determination, live-dead staining, apoptosis/necrosis, mitochondrial activity, surface markers for cell identity and characterization, determination of cell activation (immune cells, pro / anti-inflammatory), identity and function detection of transcription factors, as well as differentiation status (pluripotency, foregut endoderm, lung, etc.), detection of transgene expression in transfections and transductions (mRNA, plasmids, viral vectors), kit-based determination of cytokine concentrations
• Reference project
  • Comparison of two lab-scale protocols for enhanced mRNA-based CAR-T cell generation and functionality
  • Optimized peptide nanofibrils as efficient transduction enhancers for in vitro and ex vivo gene transfer
  • CD44v6 specific CAR-NK cells for targeted immunotherapy of head and neck squamous cell carcinoma

• Specifications: 3 lasers (405 nm, 488 nm, 640 nm), 8 fluorescence channels, high throughput measurements possible, automated staining
• Assays: Cell count determination, live-dead staining, apoptosis/necrosis, mitochondrial activity, surface markers for cell identity and characterization, surface markers + intracellular for determination of activation (immune cells, pro/anti-inflammatory), detection of transcription factors as proof of identity and function, as well as differentiation status (pluripotency, foregut endoderm, lung, etc), detection of transgene expression in transfections and transductions (mRNA, plasmids, viral vectors)
• Reference project
  • Efficient Redirection of NK Cells by Genetic Modification with Chemokine Receptors CCR4 and CCR2B
  • CD44v6 specific CAR-NK cells for targeted immunotherapy of head and neck squamous cell carcinoma

• Specifications: 2 fluorescence channels from violet (405 nm) laser, 5 from blue (488 nm) laser and 3 from red (638 nm) laser, photodiodes (forward and side scatter), photomultiplier tubes (fluorescence), high-throughput measurements possible, cell number determination possible
• Examples of assays: including immune status human blood; NK cells; regulatory T cells (Tregs); proliferation assays.