Hexagonal Boron Nitride Spacers for Fluorescence Imaging of Biomolecules

Fluorescence imaging is an invaluable tool to investigate biomolecular dynamics, mechanics, and interactions in aqueous environments. Two-dimensional materials offer large-area, atomically smooth surfaces for wide-field biomolecule imaging. Despite the success of graphene for on-chip biosensing and biomolecule manipulation, its strong...

Localization microscopy of constrained fluorescent molecules: Pushing towards Ångström-scale resolution through cryogenics

<i>Localization microscopy has circumvented the diffraction limit by sequentially</i> imaging individual light emitting molecules at a time. The position of these individual molecules can be determined and a super-resolution reconstruction is made with improved resolution. Normally freely rotating emitters are used such that the point spread...

AutoStepfinder: A fast and automated step detection method for single-molecule analysis

Single-molecule techniques allow the visualization of the molecular dynamics of nucleic acids and proteins with high spatiotemporal resolution. Valuable kinetic information of biomolecules can be obtained when the discrete states within single-molecule time trajectories are determined. Here, we present a fast, automated, and bias-free step...

An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation

Halide assays are important for the study of enzymatic dehalogenation, a topic of great industrial and scientific importance. Here we describe the development of a very sensitive halide assay that can detect less than a picomole of bromide ions, making it very useful for quantifying enzymatic dehalogenation products. Halides are oxidised...

Physical and Computational Approaches to Aberration Correction In Fluorescence Microscopy

The goal of this thesis, called Physical and Computational Approaches to Aberration Correction In Fluorescence Microscopy, concerns itself with the development of new techniques to control adaptive fluorescence microscopes, so that they can adapt and image with increased resolution, contrast and speed inside complex three-dimensional biological...

DNA sequence encodes the position of DNA supercoils

The three-dimensional organization of DNA is increasingly understood to play a decisive role in vital cellular processes. Many studies focus on the role of DNA-packaging proteins, crowding, and confinement in arranging chromatin, but structural information might also be directly encoded in bare DNA itself. Here, we visualize plectonemes ...

Intercalation-based single-molecule fluorescence assay to study DNA supercoil dynamics

DNA supercoiling crucially affects cellular processes such as DNA replication, gene expression, and chromatin organization. However, mechanistic understanding of DNA supercoiling and the related DNA-processing enzymes has remained limited, mainly due to the lack of convenient experimental tools to probe these phenomena. Here, we report a...

Photon-Counting Arrays for Time-Resolved Imaging

The paper presents a camera comprising 512 × 128 pixels capable of single-photon detection and gating with a maximum frame rate of 156 kfps. The photon capture is performed through a gated single-photon avalanche diode that generates a digital pulse upon photon detection and through a digital one-bit counter. Gray levels are obtained through...

Curved Amphipathic Oligothiophenes: Curvature directed S.A. with applications in energy transfer systems and membrane curvature detection in living cells

We have designed and synthesised a new class of molecules which self-assemble based on hydrophobic interactions with an additional driving force for influencing the overall aggregate morphology, namely, molecular curvature. The structure is build from subunits which are substituted with either hydrophilic or hydrophobic substituents. In water,...

Aerosol MALDI mass spectrometry for bioaerosol analysis

In the thesis Aerosol MALDI mass spectrometry for bioaerosol analysis is described how the aerosol mass spectrometer of the TU Delft has been further developed for the on-line analysis of bioaerosols. Due to the implemented improvements mass spectra with high resolution and a high mass range can be obtained from single protein containing aerosol...

Spatially periodic liquid crystal director field appearing in a photonic crystal template pectroscopy

Active tuning of photonic crystals can be achieved by filling the porous structures with liquid crystals. Here, the director field in macropores was studied by fluorescence confocal polarizing microscopy. For this purpose, the photonic crystal was infiltrated with a glass-forming liquid crystalline polymer, the sample was cooled below the glass...

Radiation-Induced Polymerization Monitored with Fluorogenic Molecular Probes

Each year over one billion pounds of acrylic-based polymeric products are produced world wide. Such products include windows in aircraft, lenses in eyeglasses and CD players, coatings on parquet flooring and various architectural structures such as skylights and domes. Often these products are made by injecting monomer or monomer solutions into...