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Zero-Mode Waveguide Nanowells for Single-Molecule Detection in Living Cells

Yang, Sora (author), Klughammer, N. (author), Barth, A. (author), Tanenbaum, M. (author), Dekker, C. (author)

Single-molecule fluorescence imaging experiments generally require sub-nanomolar protein concentrations to isolate single protein molecules, which makes such experiments challenging in live cells due to high intracellular protein concentrations. Here, we show that single-molecule observations can be achieved in live cells through a drastic...

journal article 2023

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Supercoiling-dependent DNA binding: quantitative modeling and applications to bulk and single-molecule experiments

Kolbeck, Pauline J. (author), Tišma, M. (author), Analikwu, B.T. (author), Vanderlinden, Willem (author), Dekker, C. (author), Lipfert, Jan (author)

DNA stores our genetic information and is ubiquitous in applications, where it interacts with binding partners ranging from small molecules to large macromolecular complexes. Binding is modulated by mechanical strains in the molecule and can change local DNA structure. Frequently, DNA occurs in closed topological forms where topology and...

journal article 2023

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Nucleotide binding halts diffusion of the eukaryotic replicative helicase during activation

Ramirez Montero, D.F. (author), Sanchez González, H. (author), van Veen, E.N.W. (author), van Laar, T. (author), Solano Hermosilla, B.P. (author), Diffley, John F.X. (author), Dekker, N.H. (author)

The eukaryotic replicative helicase CMG centrally orchestrates the replisome and leads the way at the front of replication forks. Understanding the motion of CMG on the DNA is therefore key to our understanding of DNA replication. In vivo, CMG is assembled and activated through a cell-cycle-regulated mechanism involving 36 polypeptides that...

journal article 2023

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Microwell-enhanced optical rapid antibiotic susceptibility testing of single bacteria

Roslon, I.E. (author), Japaridze, A. (author), Rodenhuis, Stef (author), Hamoen, Lieke (author), Ghatkesar, M.K. (author), Steeneken, P.G. (author), Dekker, C. (author), Alijani, F. (author)

Bacteria that are resistant to antibiotics present an increasing burden on healthcare. To address this emerging crisis, novel rapid antibiotic susceptibility testing (AST) methods are eagerly needed. Here, we present an optical AST technique that can determine the bacterial viability within 1 h down to a resolution of single bacteria. The...

journal article 2023

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De novo fabrication of custom-sequence plasmids for the synthesis of long DNA constructs with extrahelical features

Ramirez Montero, D.F. (author), Liu, Z. (author), Dekker, N.H. (author)

DNA constructs for single-molecule experiments often require specific sequences and/or extrahelical/noncanonical structures to study DNA-processing mechanisms. The precise introduction of such structures requires extensive control of the sequence of the initial DNA substrate. A commonly used substrate in the synthesis of DNA constructs is...

journal article 2023

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Identifying energy model fingerprints in mitigation scenarios

Dekker, Mark M. (author), Daioglou, Vassilis (author), Pietzcker, Robert (author), Rodrigues, Renato (author), de Boer, Harmen Sytze (author), Dalla Longa, Francesco (author), Drouet, Laurent (author), Emmerling, Johannes (author), Fattahi, Amir (author), Fotiou, Theofano (author), Fragkos, Panagiotis (author), Fricko, Oliver (author), Gusheva, E. (author), Harmsen, Mathijs (author), Huppmann, Daniel (author), Kannavou, Maria (author), Krey, Volker (author), Lombardi, F. (author), Luderer, Gunnar (author), Pfenninger, Stefan (author), Tsiropoulos, Ioannis (author), Zakeri, Behnam (author), van der Zwaan, Bob (author), Usher, Will (author), van Vuuren, Detlef (author)

Energy models are used to study emissions mitigation pathways, such as those compatible with the Paris Agreement goals. These models vary in structure, objectives, parameterization and level of detail, yielding differences in the computed energy and climate policy scenarios. To study model differences, diagnostic indicators are common practice...

journal article 2023

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Electrostatic control of the proximity effect in the bulk of semiconductor-superconductor hybrids

van Loo, N. (author), Mazur, G.P. (author), Dvir, T. (author), Wang, Guanzhong (author), Dekker, R.C. (author), Lemang, M.F. (author), Sfiligoj, C. (author), Bordin, A. (author), van Driel, D. (author), Kouwenhoven, Leo P. (author)

The proximity effect in semiconductor-superconductor nanowires is expected to generate an induced gap in the semiconductor. The magnitude of this induced gap, together with the semiconductor properties like spin-orbit coupling and g-factor, depends on the coupling between the materials. It is predicted that this coupling can be adjusted...

journal article 2023

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Detection of phosphorylation post-translational modifications along single peptides with nanopores

Nova, I.C. (author), Ritmejeris, J. (author), Brinkerhoff, H.D. (author), Koenig, T.J.R. (author), Gundlach, Jens H. (author), Dekker, C. (author)

Current methods to detect post-translational modifications of proteins, such as phosphate groups, cannot measure single molecules or differentiate between closely spaced phosphorylation sites. We detect post-translational modifications at the single-molecule level on immunopeptide sequences with cancer-associated phosphate variants by...

journal article 2023

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Three-dimensional localization microscopy with increased axial precision through TIRF angle modulation

Fan, D. (author), Cnossen, J.P. (author), Hung, S. (author), Kromm, D. (author), Dekker, N.H. (author), Verbiest, G.J. (author), Smith, C.S. (author)

To better understand the interactions between biological molecules, a high optical resolution in all three dimensions is crucial. The intrinsically lower axial resolution of microscopes however, is a limiting factor in fluorescence imaging, correspondingly in fluorescence based single molecule localization microscopy (SMLM). Here, we present...

journal article 2023

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Looping the Genome with SMC Complexes

Kim, Eugene (author), Barth, R. (author), Dekker, C. (author)

SMC (structural maintenance of chromosomes) protein complexes are an evolutionarily conserved family of motor proteins that hold sister chromatids together and fold genomes throughout the cell cycle by DNA loop extrusion. These complexes play a key role in a variety of functions in the packaging and regulation of chromosomes, and they have...

review 2023

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Diameter Dependence of Transport through Nuclear Pore Complex Mimics Studied Using Optical Nanopores

Klughammer, N. (author), Barth, A. (author), Dekker, Maurice (author), Fragasso, A. (author), Onck, Patrick R. (author)

The nuclear pore complex (NPC) regulates the selective transport of large biomolecules through the nuclear envelope. As a model system for nuclear transport, we construct NPC mimics by functionalizing the pore walls of freestanding palladium zero-mode waveguides with the FG-nucleoporin Nsp1. This approach enables the measurement of single...

journal article 2023

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Dynamin A as a one-component division machinery for synthetic cells

De Franceschi, N. (author), Barth, R. (author), Meindlhumer, S. (author), Fragasso, A. (author), Dekker, C. (author)

Membrane abscission, the final cut of the last connection between emerging daughter cells, is an indispensable event in the last stage of cell division and in other cellular processes such as endocytosis, virus release or bacterial sporulation. However, its mechanism remains poorly understood, impeding its application as a cell-division...

journal article 2023

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Unsteady flow behaviour of multi-rotors in ground proximity

Dekker, H.N.J. (author), Baars, W.J. (author), Scarano, F. (author), Tuinstra, Marthijn (author), Ragni, D. (author)

The unsteady flow behaviour of two side-by-side rotors in ground proximity is experimentally investigated. The rotors induce a velocity distribution interacting with the ground causing the radial expansion of the rotor wakes. In between the rotors, an interaction of the two wakes takes place, resulting in an upward flow similar to a fountain....

journal article 2023

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Opto-electronic feedback control of membrane potential for real-time control of action potentials

Ördög, Balázs (author), De Coster, Tim (author), Dekker, Sven O. (author), Bart, Cindy I. (author), Zhang, Juan (author), Boink, Gerard J.J. (author), Bax, Wilhelmina H. (author), Deng, Shanliang (author), den Ouden, B.L. (author), de Vries, Antoine A.F. (author), Pijnappels, Daniël A. (author)

To unlock new research possibilities by acquiring control of action potential (AP) morphologies in excitable cells, we developed an opto-electronic feedback loop-based system integrating cellular electrophysiology, real-time computing, and optogenetic approaches and applied it to monolayers of heart muscle cells. This allowed accurate...

journal article 2023

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A DNA turbine powered by a transmembrane potential across a nanopore

Shi, X. (author), Pumm, Anna Katharina (author), Maffeo, Christopher (author), Kohler, Fabian (author), Feigl, Elija (author), Zhao, W. (author), Verschueren, D.V. (author), Golestanian, Ramin (author), Aksimentiev, Aleksei (author), Dietz, Hendrik (author), Dekker, C. (author)

Rotary motors play key roles in energy transduction, from macroscale windmills to nanoscale turbines such as ATP synthase in cells. Despite our abilities to construct engines at many scales, developing functional synthetic turbines at the nanoscale has remained challenging. Here, we experimentally demonstrate rationally designed nanoscale DNA...

journal article 2023

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DAIS: The Delft Database of EEG Recordings of Dutch Articulated and Imagined Speech

Dekker, Bo (author), Schouten, A.C. (author), Scharenborg, O.E. (author)

Silent speech interfaces could enable people who lost the ability to use their voice or gestures to communicate with the external world, e.g., through decoding the person’s brain signals when imagining speech. Only a few and small databases exist that allow for the development and training of brain computer interfaces (BCIs) that can decode...

conference paper 2023

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Ultrasound Imaging with Pre-charged Collapse-Mode CMUTs

Kawasaki, S. (author), Saccher, M. (author), de Wijs, Willem-Jan (author), van den Brand, Jeroen (author), Dekker, R. (author)

Capacitive micromachined ultrasonic transducers (CMUTs) with a built-in charge layer are known as a pre-charged CMUT. In our prior work, we have shown how to model and characterize the charges inside the pre-charged collapse-mode CMUT and conducted life-time test that showed that the charges trapped inside the dielectric were stable in the order...

conference paper 2023

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A Comparative Study of Si3N4 and Al2O3 as Dielectric Materials for Pre-Charged Collapse-Mode CMUTs

Saccher, M. (author), Schaijk, Rob van (author), Kawasaki, Shinnosuke (author), Klootwijk, Johan H. (author), Rashidi, A. (author), Giagka, Vasiliki (author), Savoia, Alessandro Stuart (author), Dekker, R. (author)

Capacitive Micromachined Ultrasound Transducers (CMUTs) have many advantages compared to other ultrasonic transducer technologies, especially for implantable devices. However, they require a high bias voltage for efficient operation. To eliminate the need for an external bias voltage, a charge storage layer can be embedded in the dielectric....

conference paper 2023

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Phase Distribution Efficiency of cm-Scale Ultrasonically Powered Receivers

Saccher, M. (author), Rashidi, A. (author), Savoia, Alessandro Stuart (author), Giagka, Vasiliki (author), Dekker, R. (author)

In the domain of ultrasonically powered biomedical implants, there is an increasing interest in cm-scale ultrasonic receivers (RX). However, when a single-element transducer is used as the RX transducer, an uneven phase distribution across the RX area can significantly reduce the harvestable power. In this paper, we investigate the impact of...

conference paper 2023

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An Ultrasonically Powered System Using an AlN PMUT Receiver for Delivering Instantaneous mW-Range DC Power to Biomedical Implants

Rashidi, A. (author), Saccher, M. (author), Karuthedath, Cyril Baby (author), Sebastian, Abhilash Thanniyil (author), Savoia, Alessandro Stuart (author), Lavigne, Frederik (author), Stubbe, Frederic (author), Dekker, R. (author), Giagka, Vasiliki (author)

Aluminum Nitride (AlN) Piezoelectric Micromachined Ultrasonic Transducers (PMUTs) are gaining interest for biomedical implant power due to biocompatibility and lowtemperature processing. However, due to the low piezoelectric coefficient of AlN PMUTs, storage capacitors are often used to accumulate ultrasonic power transferred over an extended...

conference paper 2023