A5006700967- Monoclonal and Polyclonal Antibodies Research
- Advanced Fluorescence Microscopy Techniques
- Advanced Biosensing Techniques and Applications
- Glycosylation and Glycoproteins Research
- Advanced Electron Microscopy Techniques and Applications
- Cell Image Analysis Techniques
- Cellular transport and secretion
- Galectins and Cancer Biology
- Single-cell and spatial transcriptomics
- Advanced biosensing and bioanalysis techniques
- Immunotherapy and Immune Responses
- Luminescence and Fluorescent Materials
- Ion-surface interactions and analysis
- T-cell and B-cell Immunology
- RNA and protein synthesis mechanisms
- Lipid Membrane Structure and Behavior
- Near-Field Optical Microscopy
Universitätsmedizin Göttingen
2018-2024
University of Göttingen
2018-2024
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)...
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...
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.
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...
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 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...
Imaging of living synapses has relied for over two decades on the overexpression synaptic proteins fused to fluorescent reporters. This strategy alters stoichiometry components and ultimately affects synapse physiology. To overcome these limitations, here a nanobody is presented that binds calcium sensor synaptotagmin-1 (NbSyt1). functions as an intrabody (iNbSyt1) in neurons minimally invasive, leaving transmission almost unaffected, suggested by crystal structure NbSyt1 bound...
SUMMARY To fully understand biological processes and functions, it is necessary to reveal the molecular heterogeneity of cells even subcellular assemblies by gaining access location interaction all biomolecules. The study protein arrangements has seen significant advancements through super-resolution microscopy, but such methods are still far from reaching multiplexing capacity spatial proteomics. Here, we introduce Secondary label-based Unlimited Multiplexed DNA-PAINT (SUM-PAINT), a...
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 novel, rationally designed tag that serves an exceptionally broad spectrum of applications in life sciences while outperforming established like HA, FLAG myc tags. The ALFA-tag forms small and stable α-helix functional irrespective its position on target protein prokaryotic eukaryotic hosts. We developed nanobody...
Abstract Imaging of living synapses has relied for over two decades on the overexpression synaptic proteins fused to fluorescent reporters. This strategy changes stoichiometry components and ultimately affects synapse physiology. To overcome these limitations, here we introduce a nanobody that binds calcium sensor synaptotagmin-1 (NbSyt1). functions in neurons as an intrabody (iNbSyt1) is minimally invasive, leaving transmission almost unaffected, demonstrated by crystal structure NbSyt1...
Abstract DNA-PAINT is a rapidly developing fluorescence super-resolution technique which allows for reaching spatial resolutions below 10 nm. It also enables the imaging of multiple targets in same sample. However, using 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 nanobodies minimize linkage error ~4 We...
2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) is an abundant constituent of central nervous system non-compact myelin, frequently used as a marker antigen for myelinating cells. The catalytic activity CNPase, the 3'-hydrolysis nucleotides, well characterised
Abstract 2′,3′‐Cyclic nucleotide 3′‐phosphodiesterase (CNPase) is an abundant constituent of central nervous system non‐compact myelin, and its loss in mice humans causes neurodegeneration. Additionally, CNPase frequently used as a marker antigen for myelinating cells. The catalytic activity CNPase, the 3′‐hydrolysis 2′,3′‐cyclic nucleotides, well characterised vitro, but vivo function remains unclear. interacts with actin cytoskeleton to counteract developmental closure cytoplasmic channels...
Abstract The most common procedure to reveal the location of specific (sub)cellular elements in biological samples is via immunostaining followed by optical imaging. This typically performed with target-specific primary antibodies (1.Abs), which are revealed fluorophore-conjugated secondary (2.Abs). However, at high resolution this methodology can induce a series artifacts due large size antibodies, their bivalency, and polyclonality. Here we use STED DNA-PAINT super-resolution microscopy or...
Correction for ‘Circumvention of common labelling artefacts using secondary nanobodies’ by Shama Sograte-Idrissi <italic>et al.</italic>, <italic>Nanoscale</italic>, 2020, <bold>12</bold>, 10226–10239, DOI: 10.1039/D0NR00227E.
We developed here an iodine-containing probe that can be used to identify the molecules of interest in secondary ion mass spectrometry (SIMS) by simple immunolabelling procedures. The immunolabelled iodine was readily combined with previously-developed SIMS probes carrying fluorine, generate dual-channel data. This should provide a useful complement currently available probes, thus expanding scope this technology.