Multi-color, quantitative single-molecule localization microscopy (qSMLM) based on kinetic analysis of fluorophore blinking cycles

• Applicant: Professor Dr. Mike Heilemann
• Subject Area: Biophysics ; Physical Chemistry of Molecules, Liquids and Interfaces, Biophysical Chemistry
• Term: from 2016 to 2020
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 316699192

• Final Report Year: 2020

Final Report Abstract

The aim of the project was to develop quantitative single-molecule super-resolution microscopy by extracting kinetic information from single fluorophores, and to establish this technology for cell biology applications. All of the objectives of the project were achieved. The technology of qSMLM has attracted considerable response in the research community, with various research groups now using this technology. It has also become a robust standard technology in my research group.

Publications

• Molecule Counts in Localization Microscopy with Organic Fluorophores. Chemphyschem. 2017 Apr 19;18(8):942-948
  Karathanasis C, Fricke F, Hummer G, Heilemann M
  (See online at https://doi.org/10.1002/cphc.201601425)
• A toolbox for multiplexed super-resolution imaging of the E. coli nucleoid and membrane using novel PAINT labels. Sci Rep. 2018 Oct 3;8(1):14768
  Spahn CK, Glaesmann M, Grimm JB, Ayala AX, Lavis LD, Heilemann M
  (See online at https://doi.org/10.1038/s41598-018-33052-3)
• Protein-Specific, Multicolor and 3D STED Imaging in Cells with DNA-Labeled Antibodies. Angew Chem Int Ed Engl. 2019 Dec 19;58(52):18835-18838
  Spahn C, Hurter F, Glaesmann M, Karathanasis C, Lampe M, Heilemann M
  (See online at https://doi.org/10.1002/anie.201910115)
• Synthetic and genetic dimers as quantification ruler for single-molecule counting with PALM. Mol Biol Cell. 2019 Jun 1;30(12):1369-1376
  Baldering TN, Dietz MS, Gatterdam K, Karathanasis C, Wieneke R, Tampé R, Heilemann M
  (See online at https://doi.org/10.1091/mbc.E18-10-0661)
• Quantitative single-molecule imaging of TNFR1 reveals zafirlukast as antagonist of TNFR1 clustering and TNFα-induced NF-ĸB signaling. J Leukoc Biol. 2020 May 13
  Weinelt N, Karathanasis C, Smith S, Medler J, Malkusch S, Fulda S, Wajant H, Heilemann M, van Wijk SJL
  (See online at https://doi.org/10.1002/JLB.2AB0420-572RR)
• Single-molecule analysis reveals agonist-specific dimer formation of µ-opioid receptors. Nat Chem Biol. 2020 Jun 15
  Möller J, Isbilir A, Sungkaworn T, Osberg B, Karathanasis C, Sunkara V, Grushevskyi EO, Bock A, Annibale P, Heilemann M, Schütte C, Lohse MJ
  (See online at https://doi.org/10.1038/s41589-020-0566-1)
• Single-molecule imaging reveals the oligomeric state of functional TNFα-induced plasma membrane TNFR1 clusters in cells. Sci Signal. 2020 Jan 14;13(614):eaax5647
  Karathanasis C, Medler J, Fricke F, Smith S, Malkusch S, Widera D, Fulda S, Wajant H, van Wijk SJL, Dikic I, Heilemann M
  (See online at https://doi.org/10.1126/scisignal.aax5647)