@phdthesis{Nordblom2023, author = {Nordblom, Noah Frieder}, title = {Synthese und Evaluation von Gephyrinsonden f{\"u}r hochaufl{\"o}sende Mikroskopieverfahren}, doi = {10.25972/OPUS-30230}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-302300}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2023}, abstract = {This decade saw the development of new high-end light microscopy approaches. These technologies are increasingly used to expand our understanding of cellular function and the molecular mechanisms of life and disease. The precision of state-of-the-art super resolution microscopy is limited by the properties of the applied fluorescent label. Here I describe the synthesis and evaluation of new functional fluorescent probes that specifically stain gephyrin, universal marker of the neuronal inhibitory post-synapse. Selected probe precursor peptides were synthesised using solid phase peptide synthesis and conjugated with selected super resolution capable fluorescent dyes. Identity and purity were defined using chromatography and mass spectrometric methods. To probe the target specificity of the resulting probe variants in cellular context, a high-throughput assay was established. The established semi-automated and parallel workflow was used for the evaluation of three selected probes by defining their co-localization with the expressed fluorescent target protein. My work provided NN1Dc and established the probe as a visualisation tool for essentially background-free visualisation of the synaptic marker protein gephyrin in a cellular context. Furthermore, NN1DA became part of a toolbox for studying the inhibitory synapse ultrastructure and brain connectivity and turned out useful for the development of a label-free, high-throughput protein interaction quantification assay.}, subject = {Fluoreszenzmikroskopie}, language = {en} }