A5091197463- Advanced Fluorescence Microscopy Techniques
- Monoclonal and Polyclonal Antibodies Research
- Advanced Electron Microscopy Techniques and Applications
- Cellular transport and secretion
- Advanced biosensing and bioanalysis techniques
- Advanced Biosensing Techniques and Applications
- Lipid Membrane Structure and Behavior
- Cell Image Analysis Techniques
- Neuroscience and Neuropharmacology Research
- Parkinson's Disease Mechanisms and Treatments
- Ion-surface interactions and analysis
- Near-Field Optical Microscopy
- T-cell and B-cell Immunology
- RNA Interference and Gene Delivery
- Glycosylation and Glycoproteins Research
- Photoreceptor and optogenetics research
- Carbon Nanotubes in Composites
- Mass Spectrometry Techniques and Applications
- Molecular Junctions and Nanostructures
- Immune Cell Function and Interaction
- Hearing, Cochlea, Tinnitus, Genetics
- DNA and Nucleic Acid Chemistry
- Neurological disorders and treatments
- Receptor Mechanisms and Signaling
- RNA and protein synthesis mechanisms
University of Göttingen
2016-2025
Universitätsmedizin Göttingen
2015-2025
Immatics Biotechnologies (Germany)
2023-2024
Nanoscale Microscopy and Molecular Physiology of the Brain Cluster of Excellence 171 — DFG Research Center 103
2008-2019
Institute of Microbiology
2018
Universidad Santo Tomás
2018
European Neuroscience Institute Göttingen
2010-2015
Deutsche Forschungsgemeinschaft
2010-2012
University of Chile
2004
Abstract Specialized epitope tags are widely used for detecting, manipulating or purifying proteins, but often their versatility is limited. Here, we introduce the ALFA-tag, a rationally designed tag that serves remarkably broad spectrum of applications in life sciences while outperforming established like HA-, FLAG®- myc-tag. The ALFA-tag forms small and stable α-helix functional irrespective its position on target protein prokaryotic eukaryotic hosts. We characterize nanobody (NbALFA)...
The turnover of brain proteins is critical for organism survival, and its perturbations are linked to pathology. Nevertheless, protein lifetimes have been difficult obtain in vivo. They readily measured vitro by feeding cells with isotopically labeled amino acids, followed mass spectrometry analyses. In vivo generated from at least two sources: acids the diet, non-labeled degradation pre-existing proteins. This renders measurements difficult. Here we solved this problem rigorously a workflow...
Microtubules are hollow biopolymers of 25-nm diameter and key constituents the cytoskeleton. In neurons, microtubules organized differently between axons dendrites, but their precise organization in different compartments is not completely understood. Super-resolution microscopy techniques can detect specific structures at an increased resolution, narrow spacing neuronal poses challenges because most existing labelling strategies increase effective microtubule by 20-40 nm will thereby blend...
To understand biological processes, it is necessary to reveal the molecular heterogeneity of cells by gaining access location and interaction all biomolecules. Significant advances were achieved super-resolution microscopy, but such methods are still far from reaching multiplexing capacity proteomics. Here, we introduce secondary label-based unlimited multiplexed DNA-PAINT (SUM-PAINT), a high-throughput imaging method that capable achieving virtually at better than 15 nm resolution. Using...
Neurotransmitter release is achieved through the fusion of synaptic vesicles with neuronal plasma membrane (exocytosis). Vesicles are then retrieved from (endocytosis). It was hypothesized more than 3 decades ago that endosomes participate in vesicle recycling, constituting a slow endocytosis pathway required especially after prolonged stimulation. This recycling model predicts newly endocytosed fuse an endosome, which sorts (organizes) molecules and buds exocytosis-competent vesicles. We...
The recent 2014 Nobel Prize in chemistry honored an era of discoveries and technical advancements the field super‐resolution microscopy. However, applications diffraction‐unlimited imaging biology have a long road ahead persistently engage scientists with new challenges. Some bottlenecks that restrain dissemination techniques are tangible, include limited performance affinity probes yet not capillary diffusion setups. Likewise, microscopy has introduced paradigms design projects require...
The transferrin receptor, CD71, is an attractive target for drug development because of its high expression on a number cancer cell lines and the blood brain barrier. To generate serum-stabilized aptamers that recognize human we have modified traditional aptamer selection protocol by employing functional step enriches RNA molecules which bind receptor are internalized cells. Selected were specific rapidly endocytosed cells shared common core structure. A minimized variant was found to...
Secondary nanobodies can minimize probe-induced clusters artefacts. Their small size also allows fast sample penetration, and their monovalent binding enables multiplex staining using primaries from the same species.
Abstract Single‐walled carbon nanotubes (SWCNTs) are a 1D nanomaterial that shows fluorescence in the near‐infrared (NIR, >800 nm). In past, covalent chemistry was less explored to functionalize SWCNTs as it impairs NIR emission. However, certain sp 3 defects (quantum defects) lattice have emerged preserve and even introduce new, red‐shifted emission peak. Here, we report on quantum defects, introduced using light‐driven diazonium chemistry, serve anchor points for peptides proteins. We...
DNA point accumulation for imaging in nanoscale topography (DNA-PAINT) is a powerful super-resolution technique highly suitable multi-target (multiplexing) bio-imaging. However, multiplexed of cells still challenging due to the dense and sticky environment inside cell. Here, we combine fluorescence lifetime microscopy (FLIM) with DNA-PAINT use information as multiplexing parameter targets identification. In contrast Exchange-PAINT, PAINT (FL-PAINT) can image multiple simultaneously does not...
Abstract Fluorescence imaging is one of the most versatile and widely-used tools in biology 1 . Although techniques to overcome diffraction barrier were introduced more than two decades ago, nominal attainable resolution kept improving 2, 3 , fluorescence microscopy still fails image morphology single proteins or small molecular complexes, either purified a cellular context 4, 5 Here we report solution this problem, form o ne-step n anoscale e xpansion (ONE) microscopy. We combined 10-fold...
We present an in-depth analysis of selected CASP15 targets, focusing on their biological and functional significance. The authors the structures identify discuss key protein features evaluate how effectively these aspects were captured in submitted predictions. While overall ability to predict three-dimensional continues impress, reproducing uncommon not previously observed experimental is still a challenge. Furthermore, instances with conformational flexibility large multimeric complexes...
The attainable resolution of fluorescence microscopy has reached the subnanometer range, but this technique still fails to image morphology single proteins or small molecular complexes. Here, we expand specimens at least tenfold, label them with conventional fluorophores and light microscopes, acquiring videos in which analyze fluctuations. One-step nanoscale expansion (ONE) enables visualization shapes individual membrane soluble proteins, achieving around 1-nm resolution. We show that...
Deuterium (2H) MRI is an emerging tool for noninvasive imaging. We explore the integration of 2H with deuterated multifunctional nanopolymers particle imaging (DPI). To this end, amine-terminated G5-polyamidoamine (PAMAM) dendrimers were labeled acetyl surface groups, leading to highly 2H-loaded bioparticles, making them ideal studies. The accumulation ∼5 nm PAMAM in kidneys could then be seen by high submillimeter resolution. natural abundance HDO signal provided internal concentration...
Synaptic vesicles recycle repeatedly in order to maintain synaptic transmission. We have previously proposed that upon exocytosis the vesicle components persist as clusters, which would be endocytosed whole units. It has also been diffuse into plasma membrane and are then randomly gathered new vesicles. found here while strong stimulation (releasing entire recycling pool) causes diffusion of marker synaptotagmin out boutons, moderate approximately 19% all vesicles) is followed by no...
Abstract In fluorescence microscopy, the distribution of emitting molecule number in space is usually obtained by dividing measured that a single emitter. However, brightness individual emitters may vary strongly sample or be inaccessible. Moreover, with increasing (super-) resolution, fewer molecules are found per pixel, making this approach unreliable. Here we map exploiting fact emits only photon at time. Thus, analysing simultaneous arrival multiple photons during confocal imaging, can...
DNA point accumulation for imaging in nanoscale topography (PAINT) is a rapidly developing fluorescence super-resolution technique, which allows reaching spatial resolutions below 10 nm. It also enables the of multiple targets same sample. However, using DNA-PAINT to observe cellular structures at such resolution remains challenging. Antibodies, are commonly used this purpose, lead displacement between target protein and reporting fluorophore 20–25 nm, thus limiting resolving power. Here, we...
Abstract Fluorescent nanomaterials such as single‐walled carbon nanotubes (SWCNTs) have many advantages in terms of their photophysics, but it is difficult to target them specific locations living systems. In contrast, the green fluorescent protein (GFP) has been genetically fused proteins cells and organisms. Therefore, GFP can be seen not only a fluorophore universal target/handle. Here, we report conjugation GFP‐binding nanobodies DNA‐wrapped SWCNTs. This approach combines targeting...
Abstract Stimulation of the B cell antigen receptor (BCR) triggers signaling pathways that promote differentiation cells into plasma cells. Despite pivotal function BCR in activation, organization on surface resting and antigen-activated remains unclear. Here we show, using STED super-resolution microscopy, IgM-containing BCRs exist predominantly as monomers dimers membrane cells, but form higher oligomeric clusters upon stimulation. By contrast, a chronic lymphocytic leukemia-derived forms...
Abstract Nuclear magnetic resonance (NMR) is widely applied from analytics to biomedicine although it an inherently insensitive phenomenon. Overcoming sensitivity challenges key further broaden the applicability of NMR and, for example, improve medical diagnostics. Here, we present a rapid strategy enhance signals 13 C‐labelled metabolites with para ‐hydrogen in particular, C‐pyruvate, important molecule energy metabolism. We succeeded obtain average 27 % C polarization 1‐ C‐pyruvate water...
Abstract Secondary ion mass spectrometry (SIMS) is generally used in imaging the isotopic composition of various materials. It becoming increasingly popular biology, especially for investigations cellular metabolism. However, individual proteins are difficult to identify SIMS, which limits ability this technology study compartments or protein complexes. We present a method s pecific and fluorescence labeling (SPILL), based on novel click reaction with probes. Using method, we added 19...