A5010947833- Advanced Fluorescence Microscopy Techniques
- Nuclear Structure and Function
- Advanced Electron Microscopy Techniques and Applications
- RNA Research and Splicing
- CRISPR and Genetic Engineering
- Genomics and Chromatin Dynamics
- Epigenetics and DNA Methylation
- RNA and protein synthesis mechanisms
- Cell Image Analysis Techniques
- Microtubule and mitosis dynamics
- Pluripotent Stem Cells Research
- Reproductive Biology and Fertility
- Integrated Circuits and Semiconductor Failure Analysis
- PARP inhibition in cancer therapy
- Toxin Mechanisms and Immunotoxins
- DNA Repair Mechanisms
- Calcium signaling and nucleotide metabolism
- Chromosomal and Genetic Variations
- Biotin and Related Studies
- 3D Printing in Biomedical Research
- Virus-based gene therapy research
- Molecular Biology Techniques and Applications
- Preterm Birth and Chorioamnionitis
- Dietetics, Nutrition, and Education
- Advanced Proteomics Techniques and Applications
European Molecular Biology Laboratory
2013-2021
European Molecular Biology Laboratory
2013-2020
European Molecular Biology Organization
2017-2019
European Bioinformatics Institute
2017-2019
University of Edinburgh
2007-2008
University of Auckland
2001
An open-source, real-time fitter for 3D single-molecule localization microscopy uses experimental point spread functions. This enables accurate super-resolution on standard microscopes without dedicated optics. We present a using functions (PSFs) that achieves minimal uncertainty in any microscope and is compatible with PSF engineering approach. used this method to image cellular structures attained unprecedented quality astigmatic PSFs. The compensates most optical aberrations makes broadly...
At the beginning of mammalian life, genetic material from each parent meets when fertilized egg divides. It was previously thought that a single microtubule spindle is responsible for spatially combining two genomes and then segregating them to create two-cell embryo. We used light-sheet microscopy show bipolar spindles form in zygote independently congress maternal paternal genomes. These aligned their poles before anaphase but kept parental apart during first cleavage. This assembly...
Abstract The nuclear pore complex (NPC) is one of the largest and most protein assemblies in cell and, among other functions, serves as gatekeeper nucleocytoplasmic transport. Unraveling its molecular architecture functioning has been an active research topic for decades with recent cryogenic electron microscopy super‐resolution studies advancing our understanding NPC complex. However, specific direct visualization single copies proteins thus far elusive. Herein, we combine...
The defining activity of the homeodomain protein Nanog is ability to confer cytokine-independent self-renewal upon ES (embryonic stem) cells in which it overexpressed. However, biochemical basis by achieves this function remains unknown. In present study, we show that dimerizes through a functionally critical domain. Co-immunoprecipitation molecules tagged with distinct epitopes demonstrates self-associates region every fifth residue tryptophan. vitro binding experiments establish...
Abstract Photoactivatable fluorophores are important for single-particle tracking and super-resolution microscopy. Here we present a photoactivatable fluorophore that forms bright silicon rhodamine derivative through light-dependent protonation. In contrast to other fluorophores, no caging groups required, nor there any undesired side-products released. Using this fluorophore, create probes HaloTag actin live-cell single-molecule localization microscopy experiments. The unusual mechanism of...
Nuclear pore complexes (NPCs) are large macromolecular machines that mediate the traffic between nucleus and cytoplasm. In vertebrates, each NPC consists of ∼1000 proteins, termed nucleoporins, has a mass more than 100 MDa. While pseudo-atomic static model central scaffold recently been assembled by integrating data from isolated proteins complexes, many structural components still remain elusive due to enormous size flexibility NPC. Here, we explored power three-dimensional (3D)...
To prevent genome instability, mitotic exit is delayed until all chromosomes are properly attached to the spindle by assembly checkpoint (SAC). In this study, we characterized function of ARHGEF17, identified in a genome-wide RNA interference screen for human mitosis genes. Through series quantitative imaging, biochemical, and biophysical experiments, showed that ARHGEF17 essential SAC activity, because it major targeting factor controls localization kinase Mps1 kinetochore. This mediated...
Abstract The nuclear pore complex (NPC) is one of the largest and most protein assemblies in cell and, among other functions, serves as gatekeeper nucleocytoplasmic transport. Unraveling its molecular architecture functioning has been an active research topic for decades with recent cryogenic electron microscopy super‐resolution studies advancing our understanding NPC complex. However, specific direct visualization single copies proteins thus far elusive. Herein, we combine...
Abstract Quantitative fluorescence and superresolution microscopy are often limited by insufficient data quality or artifacts. In this context, it is essential to have biologically relevant control samples benchmark optimize the of microscopes, labels imaging conditions. Here we exploit stereotypic arrangement proteins in nuclear pore complex as situ reference structures characterize performance a variety modalities. We created four genome edited cell lines which endogenously labeled...
Abstract Gene tagging with fluorescent proteins is essential to investigate the dynamic properties of cellular proteins. CRISPR/Cas9 technology a powerful tool for inserting markers into all alleles gene interest (GOI) and permits functionality physiological expression fusion protein. It evaluate such genome-edited cell lines carefully in order preclude off-target effects caused by either (i) incorrect insertion protein, (ii) perturbation protein or (iii) non-specific genomic DNA damage...
Abstract At the beginning of mammalian life genetic material from each parent meets when fertilized egg divides. It was previously thought that a single microtubule spindle is responsible to spatially combine two genomes and then segregate them create two-cell embryo. Utilizing light-sheet microscopy, we showed bipolar spindles form in zygote, independently congress maternal paternal genomes. These aligned their poles prior anaphase but kept parental apart during first cleavage. This...
SUMMARY Essential biological functions, such as mitosis, require tight coordination of hundreds proteins in space and time. Localization, timing interactions changes cellular structure are all crucial to ensure correct assembly, function regulation protein complexes 1-4 . Live cell imaging can reveal distributions dynamics but experimental theoretical challenges prevented its use produce quantitative data a model mitosis that comprehensively integrates information enables analysis the...
Abstract We present a fitter for 3D single-molecule localization of arbitrary, experimental point spread functions (PSFs) that reaches minimum uncertainty EMCCD and sCMOS cameras, achieves more than 10 5 fits/s. provide tools to robustly model PSFs correct depth induced aberrations, which allowed us achieve an unprecedented resolution with engineered astigmatic PSFs, acquire high quality superresolution images even on standard microscopes without optics.