Genetic code expansion and bioorthogonal click labeling for super-resolution imaging of receptors with molecular resolution (A04)

• Subject Area: Biophysics ; Analytical Chemistry ; Bioinformatics and Theoretical Biology ; Organic Molecular Chemistry - Synthesis and Characterisation ; Structural Biology
• Term: from 2015 to 2020
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 258780946

The size of fluorescent probes will become the main limiting factor of high-resolution receptor imaging. We use biorthogonal click chemistry to introduce noncanonical amino acids in plasma membrane receptors for specific labeling. Single-molecule lattice light-sheet microscopy is then combined with expansion microscopy to image receptors in cells and tissue slices at 5-10 nm resolution. We use convolutional networks, particle averaging and automated image analysis to quantify receptor clusters in large volumes. These methods are used to investigate the stoichiometry of G protein-coupled receptors and tumor necrosis factor receptor clusters – two unresolved questions in receptor signaling.

• DFG Programme: CRC/Transregios
• Subproject of: TRR 166: High-end light microscopy elucidates membrane receptor function - ReceptorLight
• Applicant Institution: Friedrich-Schiller-Universität Jena
• Project Heads: Professor Dr. Rainer Heintzmann, until 6/2019; Professor Dr. Philip Kollmannsberger; Professor Dr. Markus Sauer