Show simple item record

Advancements in Nanobody Applications: From Live Synaptic Imaging to Multiplexed Fluorescence Microscopy and Nanoscale Topography Mapping

dc.contributor.advisorOpazo Dávila, Luis Felipe Dr.
dc.contributor.authorMougios, Nikolaos
dc.date.accessioned2024-12-09T18:12:12Z
dc.date.available2024-12-16T00:50:10Z
dc.date.issued2024-12-09
dc.identifier.urihttp://resolver.sub.uni-goettingen.de/purl?ediss-11858/15666
dc.identifier.urihttp://dx.doi.org/10.53846/goediss-10917
dc.format.extent184de
dc.language.isoengde
dc.subject.ddc570de
dc.titleAdvancements in Nanobody Applications: From Live Synaptic Imaging to Multiplexed Fluorescence Microscopy and Nanoscale Topography Mappingde
dc.typedoctoralThesisde
dc.contributor.refereeJakobs, Stefan Prof. Dr.
dc.date.examination2024-05-13de
dc.description.abstractengFluorescence microscopy facilitates biological research by enabling straightforward visualization of fluorescently labeled proteins in live and fixed samples. Despite significant progress in microscopy techniques, including those enabling molecular-level resolution, challenges persist in the availability of small, minimally invasive affinity probes, such as nanobodies, and in the simultaneous detection of multiple targets. These challenges are particularly significant when studying small and complex subcellular compartments such as neuronal synapses. Therefore, new small molecular tools and labeling strategies compatible with modern fluorescence microscopy are essential to reveal target proteins. To address these limitations, in this work, I initially produced and characterized a novel nanobody against synaptotagmin 1 (NbSyt1) offering precise imaging of synapses without perturbing synaptic function. NbSyt1 also functions as an intrabody in living neurons, exhibiting minimal invasiveness and enabling the generation of protein-based fluorescent reporters for super- resolution imaging. Thereafter, to enable the detection of multiple targets within single samples, I developed NanoPlex, a versatile method for multiplexing based on repetitive imaging cycles facilitated by engineered nanobodies that bind selectively to primary antibodies (secondary nanobodies) and carry adaptable cleavable fluorescent groups. Finally, I utilized some of these technologies to investigate neuronal physiology and study the dynamics of trafficking proteins within living postsynaptic compartments, as well as to resolve the effect of liquid-liquid phase separation (LLPS) on the neuronal localization of several proteins in individual neurons.de
dc.contributor.coRefereeEnderlein, Jörg Prof. Dr.
dc.subject.engSingle-domain antibodiesde
dc.subject.engCleavable probesde
dc.subject.engMultiplexingde
dc.subject.engFRAPde
dc.subject.engSuper-resolution microscopyde
dc.subject.engNeuronal imagingde
dc.subject.engNeurosciencede
dc.identifier.urnurn:nbn:de:gbv:7-ediss-15666-1
dc.affiliation.instituteGöttinger Graduiertenschule für Neurowissenschaften, Biophysik und molekulare Biowissenschaften (GGNB)de
dc.subject.gokfullBiologie (PPN619462639)de
dc.description.embargoed2024-12-16de
dc.identifier.ppn191161925X
dc.identifier.orcidhttps://orcid.org/0009-0007-5712-7175de
dc.notes.confirmationsentConfirmation sent 2024-12-09T19:45:01de


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record