Methods of Investigation

Otorhinolaryngology

Head of Department:
Prof. Dr. med. Dr. h. c. Heinrich Iro

Cyto-/Genotoxicity

  • Proliferation, morphological cell analysis: (Real-time cell imaging: IncuCyte)
  • Plasma-Membrane Integrity (LDH, spectrophotometer)
  • Cell proliferation (BrdU, spectrophotometer, flow cytometer)
  • Caspase-3/7 activation (flow cytometer)
  • Metabolic activity (MTT, WST, XTT, spectrophotometer)
  • Apoptosis/Necrosis (Annexin V/ Propidium iodide staining, flow cytometer)
  • Mitochondrial-membrane-Potential (DiIC1(5), flow cytometer)
  • Cell cycle and DNA degradation (Propidium iodide-Triton, flow cytometer)
  • Calcium-Influx (Fluo-4, flow cytometer)
  • ATP-quantification (CellTiter-Glo, spectrophotometer)
  • DNA-double-strand-breaks (yH2AX, Western blot, flow cytometer)
  • DNA-damages (COMET-Assay, fluorescent microscopy)

Oxidative stress

  • Intracellular H2O2 (DCFH, flow cytometer)
  • Intracellular O2 (Hydroethidium, flow cytometer)
  • Measurement of reduced glutathione (Monobromobimane, flow cytometer)
  • Membrane-peroxidation (BODIPY 581/591 C11, flow cytometer)
  • Lipid-peroxidation (Identification of malondialdehyd using thiobarbituric acid method, spectrophotometer)

Histology

Cryo-fixation and production of thin sections
Paraffin embedding and production of thin sections

Methods of staining

  • Prussian blue
  • Haematoxilin-Eosin
  • Masson-Goldner Trichrome staining
  • Characterization of cellular components using immunofluorescent staining (e.g. DAPI, LAMP-1, F-Actin, GM 130, EGFR, Transferrin receptor CD 71)
 
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