12 results on '"Anna E C, Meijering"'
Search Results
2. Implementation of 3D Multi-Color Fluorescence Microscopy in a Quadruple Trap Optical Tweezers System
- Author
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Anna E C, Meijering, Julia A M, Bakx, Tianlong, Man, Iddo, Heller, Gijs J L, Wuite, and Erwin J G, Peterman
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Microscopy, Fluorescence ,Optical Tweezers ,Nanotechnology ,Optical Devices ,DNA - Abstract
Recent advances in the design and measurement capabilities of optical tweezers instruments, and especially the combination with multi-color fluorescence detection, have accommodated a dramatic increase in the versatility of optical trapping. Quadruple (Q)-trap optical tweezers are an excellent example of such an advance, by providing three-dimensional control over two constructs and thereby enabling for example DNA-DNA braiding. However, the implementation of fluorescence detection in such a Q-trapping system poses several challenges: (1) since typical samples span a distance in the order of tens of micrometers, it requires imaging of a large field of view, (2) in order to capture fast molecular dynamics, fast imaging with single-molecule sensitivity is desired, (3) in order to study three-dimensional objects, it could be needed to detect emission light at different axial heights while keeping the objective lens and thus the optically trapped microspheres in a fixed position. In this chapter, we describe design guidelines for a fluorescence imaging module on a Q-trap system that overcomes these challenges and provide a step-by-step description for construction and alignment of such a system. Finally, we present detailed instructions for proof-of-concept experiments that can be used to validate and highlight the capabilities of the instruments.
- Published
- 2022
3. Nonlinear mechanics of human mitotic chromosomes
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Anna E. C. Meijering, Kata Sarlós, Christian F. Nielsen, Hannes Witt, Janni Harju, Emma Kerklingh, Guus H. Haasnoot, Anna H. Bizard, Iddo Heller, Chase P. Broedersz, Ying Liu, Erwin J. G. Peterman, Ian D. Hickson, Gijs J. L. Wuite, Physics of Living Systems, LaserLaB - Molecular Biophysics, and LaserLaB - Energy
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Multidisciplinary - Abstract
In preparation for mitotic cell division, the nuclear DNA of human cells is compacted into individualized, X-shaped chromosomes1. This metamorphosis is driven mainly by the combined action of condensins and topoisomerase IIα (TOP2A)2,3, and has been observed using microscopy for over a century. Nevertheless, very little is known about the structural organization of a mitotic chromosome. Here we introduce a workflow to interrogate the organization of human chromosomes based on optical trapping and manipulation. This allows high-resolution force measurements and fluorescence visualization of native metaphase chromosomes to be conducted under tightly controlled experimental conditions. We have used this method to extensively characterize chromosome mechanics and structure. Notably, we find that under increasing mechanical load, chromosomes exhibit nonlinear stiffening behaviour, distinct from that predicted by classical polymer models4. To explain this anomalous stiffening, we introduce a hierarchical worm-like chain model that describes the chromosome as a heterogeneous assembly of nonlinear worm-like chains. Moreover, through inducible degradation of TOP2A5 specifically in mitosis, we provide evidence that TOP2A has a role in the preservation of chromosome compaction. The methods described here open the door to a wide array of investigations into the structure and dynamics of both normal and disease-associated chromosomes.
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- 2022
- Full Text
- View/download PDF
4. Nonlinear mechanics of human mitotic chromosomes
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Anna E C, Meijering, Kata, Sarlós, Christian F, Nielsen, Hannes, Witt, Janni, Harju, Emma, Kerklingh, Guus H, Haasnoot, Anna H, Bizard, Iddo, Heller, Chase P, Broedersz, Ying, Liu, Erwin J G, Peterman, Ian D, Hickson, and Gijs J L, Wuite
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Optics and Photonics ,DNA Topoisomerases, Type II ,Chromosomes, Human ,Humans ,Mitosis ,DNA ,Chromosomes - Abstract
In preparation for mitotic cell division, the nuclear DNA of human cells is compacted into individualized, X-shaped chromosomes
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- 2021
5. Publisher Correction: Nonlinear mechanics of human mitotic chromosomes
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Anna E. C. Meijering, Kata Sarlós, Christian F. Nielsen, Hannes Witt, Janni Harju, Emma Kerklingh, Guus H. Haasnoot, Anna H. Bizard, Iddo Heller, Chase P. Broedersz, Ying Liu, Erwin J. G. Peterman, Ian D. Hickson, and Gijs J. L. Wuite
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Multidisciplinary - Abstract
In the version of this article initially published, Extended Data Fig. 5 was a duplicate of Extended Data Fig. 6. The correct image is now in place in the HTML and PDF versions of the article.
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- 2022
6. Imaging unlabeled proteins on DNA with super-resolution
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Ineke Brouwer, Anna E. C. Meijering, Andreas S. Biebricher, Gijs J.L. Wuite, Iddo Heller, Erwin J.G. Peterman, Gerrit Sitters, Physics of Living Systems, and LaserLaB - Molecular Biophysics
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AcademicSubjects/SCI00010 ,Inverse ,02 engineering and technology ,Biology ,Narese/16 ,03 medical and health sciences ,chemistry.chemical_compound ,Imaging, Three-Dimensional ,Microscopy ,Genetics ,Fluorescence microscope ,Medical imaging ,Humans ,Computer Simulation ,030304 developmental biology ,0303 health sciences ,DNA ,021001 nanoscience & nanotechnology ,Photobleaching ,Superresolution ,DNA-Binding Proteins ,Microscopy, Fluorescence ,chemistry ,Temporal resolution ,Methods Online ,0210 nano-technology ,Biological system ,Monte Carlo Method ,Protein Binding - Abstract
Fluorescence microscopy is invaluable to a range of biomolecular analysis approaches. The required labeling of proteins of interest, however, can be challenging and potentially perturb biomolecular functionality as well as cause imaging artefacts and photo bleaching issues. Here, we introduce inverse (super-resolution) imaging of unlabeled proteins bound to DNA. In this new method, we use DNA-binding fluorophores that transiently label bare DNA but not protein-bound DNA. In addition to demonstrating diffraction-limited inverse imaging, we show that inverse Binding-Activated Localization Microscopy or ‘iBALM’ can resolve biomolecular features smaller than the diffraction limit. The current detection limit is estimated to lie at features between 5 and 15 nm in size. Although the current image-acquisition times preclude super-resolving fast dynamics, we show that diffraction-limited inverse imaging can reveal molecular mobility at ∼0.2 s temporal resolution and that the method works both with DNA-intercalating and non-intercalating dyes. Our experiments show that such inverse imaging approaches are valuable additions to the single-molecule toolkit that relieve potential limitations posed by labeling.
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- 2020
7. Simultaneously Capturing the Structure and Mechanical Properties of Chromosome using STED Nanoscopy and Optical Tweezers
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Anna E. C. Meijering, Tianlong Man, Kata Sarlós, Ian D. Hickson, Gijs J.L. Wuite, Iddo Heller, and Erwin J.G. Peterman
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Physics ,Optics ,Optical tweezers ,Chromosome (genetic algorithm) ,business.industry ,Biophysics ,STED microscopy ,business - Published
- 2020
8. Exposing Chromosome Architecture and Mechanics Using Optical Manipulation and Fluorescence Microscopy
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Anna E. C. Meijering, Seyda Acar, Kata Sarlós, Iddo Heller, Ying Liu, Erwin J.G. Peterman, Rahul Bhowmick, Anna H. Bizard, Gijs J.L. Wuite, Ian D. Hickson, Andrés Bueno Venegas, Physics of Living Systems, and LaserLaB - Molecular Biophysics
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Materials science ,Chromosome architecture ,Biophysics ,Fluorescence microscope - Published
- 2018
9. Super-Resolution Imaging of Unlabeled Proteins on DNA
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Iddo Heller, Erwin J.G. Peterman, Anna E. C. Meijering, Andreas S. Biebricher, Gijs J.L. Wuite, Physics of Living Systems, LaserLaB - Molecular Biophysics, and Physics and Astronomy
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Molecular interactions ,chemistry.chemical_compound ,chemistry ,Proof of concept ,Temporal resolution ,Microscopy ,Biophysics ,Nanotechnology ,Biology ,Biological system ,Superresolution ,DNA ,Visualization - Abstract
Direct visualization of DNA-protein interactions at the single-molecule level is of ever increasing importance to unravel the salient details of a wide range of DNA-associated processes. Recent emergence of super-resolution imaging has the potential to further boost the impact of such investigations. However, current imaging methods heavily rely on (fluorescent) labeling strategies, which can be challenging and potentially even interfere with the molecular interactions under scrutiny. Here, we introduce a new label-free method to image the presence and location of DNA-bound proteins with super-resolution on optically manipulated DNA. The method is based on localization microscopy of DNA-intercalating dyes that locally bind to bare DNA but not to protein-bound DNA sections, yielding an inverted image that reveals the ‘shadows’ of the proteins on the DNA: inverse binding-activated localization microscopy (iBALM). We present the proof of principle of iBALM as well as of functional variations to this method and provide experimental and theoretical data that describe the spatial and temporal resolution that can be obtained. iBALM has the potential to become a valuable addition to the single-molecule toolkit and enable direct visualization of processes that were previously not possible due to limitations posed by labeling.
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- 2017
10. Recent Advances in Biological Single-Molecule Applications of Optical Tweezers and Fluorescence Microscopy
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Wouter H. Roos, M. Hashemi Shabestari, Gijs J.L. Wuite, Erwin J.G. Peterman, Anna E. C. Meijering, Physics of Living Systems, LaserLaB - Molecular Biophysics, and Physics and Astronomy
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0301 basic medicine ,chemistry.chemical_classification ,Fluorescence microscopy ,STED super-resolution microscopy ,Single molecule ,Biomolecule ,Microfluidics ,Resolution (electron density) ,Nanotechnology ,Molecular motors ,Optical tweezers ,Quadruple optical traps ,Single Molecule Imaging ,Commercial solutions ,03 medical and health sciences ,030104 developmental biology ,chemistry ,Fluorescence microscope ,Molecular motor ,Molecule ,DNA–protein interaction ,Mechanochemistry - Abstract
Over the past two decades, single-molecule techniques have evolved into robust tools to study many fundamental biological processes. The combination of optical tweezers with fluorescence microscopy and microfluidics provides a powerful single-molecule manipulation and visualization technique that has found widespread application in biology. In this combined approach, the spatial (~ nm) and temporal (~ ms) resolution, as well as the force scale (~ pN) accessible to optical tweezers is complemented with the power of fluorescence microscopy. Thereby, it provides information on the local presence, identity, spatial dynamics, and conformational dynamics of single biomolecules. Together, these techniques allow comprehensive studies of, among others, molecular motors, protein–protein and protein–DNA interactions, biomolecular conformational changes, and mechanotransduction pathways. In this chapter, recent applications of fluorescence microscopy in combination with optical trapping are discussed. After an introductory section, we provide a description of instrumentation together with the current capabilities and limitations of the approaches. Next we summarize recent studies that applied this combination of techniques in biological systems and highlight some representative biological assays to mark the exquisite opportunities that optical tweezers combined with fluorescence microscopy provide.
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- 2017
11. DNA-Tile Structures Induce Ionic Currents through Lipid Membranes
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Kerstin Göpfrich, Ulrich F. Keyser, Thomas Zettl, Samet Kocabey, Anna E. C. Meijering, Tim Liedl, and Silvia Hernández-Ainsa
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Materials science ,Membrane lipids ,Lipid Bilayers ,Bioengineering ,Nanotechnology ,02 engineering and technology ,Gating ,Biosensing Techniques ,010402 general chemistry ,01 natural sciences ,Membrane Lipids ,DNA nanotechnology ,General Materials Science ,Lipid bilayer ,Ion channel ,Transmembrane channels ,Mechanical Engineering ,General Chemistry ,DNA ,021001 nanoscience & nanotechnology ,Condensed Matter Physics ,0104 chemical sciences ,Nanostructures ,Membrane ,Biophysics ,Nucleic Acid Conformation ,Self-assembly ,0210 nano-technology - Abstract
Self-assembled DNA nanostructures have been used to create man-made transmembrane channels in lipid bilayers. Here, we present a DNA-tile structure with a nominal subnanometer channel and cholesterol-tags for membrane anchoring. With an outer diameter of 5 nm and a molecular weight of 45 kDa, the dimensions of our synthetic nanostructure are comparable to biological ion channels. Because of its simple design, the structure self-assembles within a minute, making its creation scalable for applications in biology. Ionic current recordings demonstrate that the tile structures enable ion conduction through lipid bilayers and show gating and voltage-switching behavior. By demonstrating the design of DNA-based membrane channels with openings much smaller than that of the archetypical six-helix bundle, our work showcases their versatility inspired by the rich diversity of natural membrane components.
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- 2015
12. Collecting optical coherence elastography depth profiles with a micromachined cantilever probe
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Johannes F. de Boer, Davide Iannuzzi, Mattijs de Groot, Anna E. C. Meijering, Jianhua Mo, D.C. Chavan, Biophotonics and Medical Imaging, Physics of Living Systems, Student Lab and Education, and LaserLaB - Biophotonics and Microscopy
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Cantilever ,Optical fiber ,Materials science ,medicine.diagnostic_test ,business.industry ,02 engineering and technology ,021001 nanoscience & nanotechnology ,01 natural sciences ,Atomic and Molecular Physics, and Optics ,law.invention ,010309 optics ,Interferometry ,Optics ,Optical coherence tomography ,law ,Deflection (engineering) ,Indentation ,0103 physical sciences ,medicine ,Elastography ,0210 nano-technology ,business ,Optomechanics - Abstract
We present an experimental setup that combines optical coherence elastography depth sensing with atomic force microscope indentation. The instrument relies on a miniaturized cantilever probe that compresses a sample with a small footprint force and simultaneously collects an optical coherence tomography (OCT) depth profile underneath the indenting point. The deflection of the cantilever can be monitored via optical fiber interferometry with a resolution of 2 nm. The OCT readout then provides depth profiles of the subsurface layer deformation with 15 nm resolution and depth range of a few millimeters. © 2013 Optical Society of America.
- Published
- 2013
- Full Text
- View/download PDF
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