A new twist on PIFE: photoisomerisation-related fluorescence enhancement

PIFE was first used as an acronym for protein-induced fluorescence enhancement, which refers to the increase in fluorescence observed upon the interaction of a fluorophore, such as a cyanine, with a protein. This fluorescence enhancement is due to changes in the rate ofcis/transphotoisomerisation. It is clear now that this mechanism is...

Exploring molecular biology in sequence space: The road to next-generation single-molecule biophysics

Next-generation sequencing techniques have led to a new quantitative dimension in the biological sciences. In particular, integrating sequencing techniques with biophysical tools allows sequence-dependent mechanistic studies. Using the millions of DNA clusters that are generated during sequencing to perform high-throughput binding affinity...

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...

Advancing single-molecule instrumentation through nanoscale optics, fabrication, and surface functionalization

This thesis describes developments in the single-molecule instrumentation, in particular optical torque wrench and DNA nanocurtains, with the goal of employing these techniques in studies of biomolecules and biomotors. Importantly, the use of single-crystal rutile titanium dioxide nanocylinders is suggested for optical torque wrench, enabling...

Heat shock proteins studied by optical tweezers

Since the first protein structure reported, the understanding of how proteins fold so fast and efficiently into such peculiar shapes has quickly become one of the major problems of biology. Protein folding in the living cell features a new level of complexity due to highly crowded cellular environment. In order to overcome this problem, cells...