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
