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Jörg Enderlein

Publications and source records attributed to Jörg Enderlein.

10 recordsLinked to original sources

Mapping complex optical light field distributions with single fluorescent molecules

Single fluorescent molecules, behaving as ideal electric dipole emitters, are powerful nanoscopic probes of complex optical fields. Here, this property is exploited to precisely map the polarization and vectorial structure of tightly focused laser beams, utilizing both linear and circular polarization states. The resulting three-dimensional fluorescence excitation maps strikingly reveal the intrinsic chiral and non-chiral structure of the light field, in perfect quantitative agreement with a full vectorial wave-optical model. This precise correspondence not only enables the reliable determination of absolute molecular orientations but also allows for the accurate characterization of the field's properties. These results fundamentally advance our understanding of light-matter interaction at the single-molecule level and open new avenues for characterizing complex light fields, with broad applications in super-resolution microscopy and nanophotonics.

physics.optics↗

Fluorescence Microscopy: a statistics-optics perspective

Fundamental properties of light unavoidably impose features on images collected using fluorescence microscopes. Modeling these features is ever more important in quantitatively interpreting microscopy images collected at scales on par or smaller than light's wavelength. Here we review the optics responsible for generating fluorescent images, fluorophore properties, microscopy modalities leveraging properties of both light and fluorophores, in addition to the necessarily probabilistic modeling tools imposed by the stochastic nature of light and measurement.

physics.optics↗

Graphene-Induced Energy Transfer for Quantitative Membrane Biophysics at Sub-Nanometer Resolution

Graphene-induced energy transfer (GIET) is a recently developed fluorescence-spectroscopic technique that achieves sub-nanometric optical localization of fluorophores along the optical axis of a microscope. GIET is based on the near-field energy transfer from an optically excited fluorescent molecule to a single sheet of graphene. It has been successfully used for estimating inter-leaflet distances of single lipid bilayers, and for investigating the membrane organization of living mitochondria. In this study, we use GIET to measure the cholesterol-induced subtle changes of membrane thickness at the nanoscale. We quantify membrane thickness variations in supported lipid bilayers (SLBs) as a function of lipid composition and increasing cholesterol content. Our findings demonstrate that GIET is an extremely sensitive tool for investigating nanometric structural changes in bio-membranes.

physics.optics↗

Measuring photophysical transition rates with fluorescence correlation spectroscopy and antibunching

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 fluorescent molecules. The antibunching measurements allow us to quantify the average excitation rate of fluorescent molecules within the confocal detection volume of the FCS measurement setup. Knowledge of this value allows us then to quantify, in an absolute manner, the intersystem crossing rate and triplet state lifetime from the microsecond temporal decay of the FCS curves. We present a theoretical analysis of the method and estimate the maximum bias caused by the averaging of all quantities (excitation rate, photophysical rates) over the confocal detection volume, and we show that this bias is smaller than 5\% in most cases. We apply the method for measuring the photophysical rate constants of the widely used dyes Rhodamine~110 and ATTO~655.

physics.chem-ph↗

Optimal transfer functions for bandwidth-limited imaging

One of the fundamental limits of classical optical microscopy is the diffraction limit of optical resolution. It results from the finite bandwidth of the optical transfer function (or OTF) of an optical microscope, which restricts the maximum spatial frequencies that are transmitted by a microscope. However, given the frequency support of the OTF, which is fully determined by the used optical hardware, an open and unsolved question is what is the optimal amplitude and phase distribution of spatial frequencies across this support that delivers the "sharpest" possible image. In this paper, we will answer this question and present a general rule how to find the optimal OTF for any given imaging system. We discuss our result in the context of optical microscopy, by considering in particular the cases of wide-field microscopy, confocal Image Scanning Microscopy (ISM), 4pi microscopy, and Structured Illumination Microscopy (SIM). Our results are important for finding optimal deconvolution algorithms for microscopy images, and we demonstrate this experimentally on the example of ISM. Our results can also serve as a guideline for designing optical systems that deliver best possible images, and they can be easily generalized to non-optical imaging such as telescopic imaging, ultrasound imaging, or magnetic resonance imaging.

physics.optics↗

Modeling Fluorescence Correlation Spectroscopy through an aberrating sphere

Fluorescence Correlation Spectroscopy (FCS) is a powerful single-molecule technique which allows for measuring motion (diffusion, flow), concentration, and molecular interaction kinetics of fluorescent molecules from picomolar to micromolar concentrations. It has found manifold applications in the physical and life sciences. Many biological/biophysical applications use FCS for measuring the motion and concentration of fluorescently labeled biomolecules in living cells and tissue. However, a correct quantitative evaluation of FCS experiments relies on the accurate knowledge of the fluorescence excitation and detection properties of the used confocal microscope. Using a bottom-up approach, we theoretically study how these properties are affected by the presence of a diffracting dielectric bead within the optical path, and how this changes the outcome of a FCS measurement. This will be important for all applications of FCS under optically non-ideal aberrating conditions.

physics.optics↗

Electric field lines of relativistically moving point charges

Generation of electromagnetic fields by moving charges is a fascinating topic where the tight connection between classical electrodynamics and special relativity becomes particularly apparent. One can gain direct insight into the fascinating structure of such fields by visualizing the electric field lines. However, the calculation of electric field lines for arbitrarily moving charges is far from trivial. Here, we derive an equation for the director that points from the retarded position of a moving charge towards a specific field line position, which allows for a simple construction of these lines. We analytically solve this equation for several special but important cases: for an arbitrary rectilinear motion, for the motion within the wiggler magnetic field of a free electron laser, and for the motion in a synchrotron.

physics.class-ph↗

Efficient solver for a special class of convection-diffusion problems

We describe an exact and highly efficient numerical algorithm for solving a special but important class of convection-diffusion equations. These equations occur in many problems in physics, chemistry, or biology, and they are usually hard to treat due to the presence of a convection term, represented by a first order derivative in the spatial co-ordinate. Our algorithm reduces the convection-diffusion equation to a pure diffusion problem within a complex-valued potential which can be solved efficiently and accurately using conventional parabolic partial differential equation solvers.

physics.comp-ph↗

An axon initial segment is required for temporal precision in action potential encoding by neuronal populations

Central neurons initiate action potentials (APs) in the axon initial segment (AIS), a compartment characterized by a high concentration of voltage-dependent ion channels and specialized cytoskeletal anchoring proteins arranged in a regular nanoscale pattern. Although the AIS was a key evolutionary innovation in neurons, the functional benefits it confers are not clear. Using a mutation of the AIS cytoskeletal protein βIV-spectrin, we here establish an in vitro model of neurons with a perturbed AIS architecture that retains nanoscale order but loses the ability to maintain a high NaV density. Combining experiments and simulations we show that a high NaV density in the AIS is not required for axonal AP initiation; it is however crucial for a high bandwidth of information encoding and AP timing precision. Our results provide the first experimental demonstration of axonal AP initiation without high axonal channel density and suggest that increasing the bandwidth of the neuronal code and hence the computational efficiency of network function was a major benefit of the evolution of the AIS.

q-bio.NC↗

Electrodynamic coupling of electric dipole emitters to a fluctuating mode density within a nano-cavity

We investigate the impact of rotational diffusion on the electrodynamic coupling of fluorescent dye molecules (oscillating electric dipoles) to a tunable planar metallic nanocavity. Fast rotational diffusion of the molecules leads to a rapidly fluctuating mode density of the electromagnetic field along the molecules' dipole axis, which significantly changes their coupling to the field as compared to the opposite limit of fixed dipole orientation. We derive a theoretical treatment of the problem and present experimental results for rhodamine 6G molecules in cavities filled with low and high viscosity liquids. The derived theory and presented experimental method is a powerful tool for determining absolute quantum yield values of fluorescence.

physics.optics↗