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A fluorescence correlation spectroscopy (FCS) system based on two independent measurement volumes is presented. The optical setup and data acquisition hardware are detailed, as well as a complete protocol to control the location, size and…

Optics · Physics 2009-08-26 P. Ferrand , M. Pianta , A. Kress , A. Aillaud , H. Rigneault , D. Marguet

In the current AFM experiments the distribution of unfolding times, P(t), is measured by applying a constant stretching force f_s from which the apparent unfolding rate is obtained. To describe the complexity of the underlying energy…

Soft Condensed Matter · Physics 2009-11-11 V. Barsegov , D. Klimov , D. Thirumalai

Determining the number of fluorescent entities that are coupled to a given molecule (DNA, protein, etc.) is a key point of numerous biological studies, especially those based on a single molecule approach. Reliable methods are important, in…

The spatial organization of enzymes often plays a crucial role in the functionality and efficiency of enzymatic pathways. To fully understand the design and operation of enzymatic pathways, it is therefore crucial to understand how the…

Subcellular Processes · Quantitative Biology 2015-06-17 Alexander Buchner , Filipe Tostevin , Florian Hinzpeter , Ulrich Gerland

Single-molecule detection at a nanometric interface in a femtomolar solution, can take weeks as the encounter rate between the diffusing molecule to be detected and the transducing nano-device is negligibly small. On the other hand, several…

By performing spectroscopic single-particle measurements at cryogenic temperatures over the course of hours we study %By performing spectroscopic longtime single-particle measurements at cryogenic temperatures we study both the spectral…

Mesoscale and Nanoscale Physics · Physics 2017-11-13 Sven-Hendrik Lohmann , Christian Strelow , Alf Mews , Tobias Kipp

Single dye molecules at cryogenic temperatures display many spectroscopic phenomena known from free atoms and are thus promising candidates for fundamental quantum optical studies. However, the existing techniques for the detection of…

Quantum Physics · Physics 2008-02-12 Gert Wrigge , Ilja Gerhardt , Jaesuk Hwang , Gert Zumofen , Vahid Sandoghdar

We adapt Fluorescence Correlation spectroscopy (FCS) formalism to the studies of the dynamics of semi-flexible polymers and derive expressions relating FCS correlation function to the longitudinal and transverse mean square displacements of…

Soft Condensed Matter · Physics 2009-11-11 Anne Bernheim-Groswasser , Roman Shusterman , Oleg Krichevsky

Physics of self-propelled objects at the nanoscale is a rapidly developing research field where recent experiments focused on motion of individual catalytic enzymes. Contrary to the experimental advancements, theoretical understanding of…

Soft Condensed Matter · Physics 2022-08-29 Artem Ryabov , Mykola Tasinkevych

Single-molecule localization microscopy allows practitioners to locate and track labeled molecules in biological systems. When extracting diffusion coefficients from the resulting trajectories, it is common practice to perform a linear fit…

Biological Physics · Physics 2024-06-19 Jakob Tómas Bullerjahn , Gerhard Hummer

The diffusion of macromolecules in cells and in complex fluids is often found to deviate from simple Fickian diffusion. One explanation offered for this behavior is that molecular crowding renders diffusion anomalous, where the mean-squared…

Soft Condensed Matter · Physics 2016-05-18 Daniel S. Banks , Charmaine Tressler , Robert D. Peters , Felix Höfling , Cécile Fradin

FRET-based approaches are a unique tool for sensing the immediate surroundings and interactions of (bio)molecules. FRET imaging and FLIM (Fluorescence Lifetime Imaging Microscopy) enable the visualization of the spatial distribution of…

We study the role of active coupling on the transport properties of homogeneously charged macromolecules in an infinitely dilute solution. An enzyme becomes actively bound to a segment of the macromolecule, exerting an electrostatic force…

Soft Condensed Matter · Physics 2024-05-24 Tapas Singha , Siao-Fong Li , Murugappan Muthukumar

We present a feature-resolved methodology to analyse the photoluminescence dynamics of single emitters using a combination of lifetime, spectral, and photon correlation analyses. By integrating conventional ensemble photon statistics…

Intensity squeezing, i.e., photon number fluctuations below the shot noise limit, is a fundamental aspect of quantum optics and has wide applications in quantum metrology. It was predicted in 1979 that the intensity squeezing could be…

We present a new method that combines fluorescence correlation spectroscopy (FCS) on the microsecond time scale with fluorescence antibunching measurements on the nanosecond time scale for measuring photophysical rate constants of…

Chemical Physics · Physics 2022-03-28 Damir Sakhapov , Ingo Gregor , Narain Karedla , Jörg Enderlein

Single molecule FRET (fluorescence resonance energy transfer) is a powerful technique for detecting real-time conformational changes and molecular interactions during biological reactions. In this review, we examine different techniques of…

Biomolecules · Quantitative Biology 2007-05-23 Ivan Rasnik , Sean A. McKinney , Taekjip Ha

Diffusion of tracer dye molecules in water confined to nanoscale is an important subject with a direct bearing on many technological applications. It is not yet clear however, if the dynamics of water in hydrophilic as well as hydrophobic…

Soft Condensed Matter · Physics 2022-01-19 Shivprasad Patil , V. J. Ajith

Numerical simulation is used to characterise double potential step chronoamperometry at a microband electrode for a simple redox process A + e- goes to B, under conditions of full support such that diffusion is the only active form of mass…

Chemical Physics · Physics 2013-10-28 Edward O. Barnes , Linhongjia Xiong , Kristopher R. Ward , Richard G. Compton

A method is proposed to pin down an unambiguous proof for single molecule surface enhanced Raman spectroscopy (SERS). The simultaneous use of two analyte molecules enables a clear confirmation of the single (or few) molecule nature of the…

Chemical Physics · Physics 2007-05-23 E. C. Le Ru , M. Meyer , P. G. Etchegoin