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The point spread function (PSF) is fundamental to any type of microscopy, most importantly so for single-molecule localization techniques, where the exact PSF shape is crucial for precise molecule localization at the nanoscale. However,…
Over the last decade, single-molecule optical microscopy has become the gold-standard approach to decipher complex molecular processes in cellular environments. [1-3] Single-molecule fluorescence microscopy has several advantages such as…
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…
In this paper we review the applicability of autofluorescent proteins for single-molecule imaging in biology. The photophysical characteristics of several mutants of the Green Fluorescent Protein (GFP) and those of DsRed are compared and…
Optical detection and spectroscopy of single molecules has become an indispensable tool in biological imaging and sensing. Its success is based on fluorescence of organic dye molecules under carefully engineered laser illumination. In this…
The movement of molecules inside living cells is a fundamental feature of biological processes. The ability to both observe and analyse the details of molecular diffusion in vivo at the single molecule and single cell level can add…
The Surface Brightness Fluctuations (SBF) Method for distance determinations of elliptical galaxies is been modeled in order to investigate the effect of the Point Spread Function (PSF). We developed a method to simulate observations of SBF…
Point Spread Function (PSF) engineering is used in single emitter localization to measure the emitter position in 3D and possibly other parameters such as the emission color or dipole orientation as well. Advanced PSF models such as spline…
Standard deviation measurements of intensity profiles of stationary single fluorescent molecules are useful for studying axial localization, molecular orientation, and a fluorescence imaging system's spatial resolution. Here we report on…
The knowledge of the exact structure of the optical system PSF enables a high-quality image reconstruction in fluorescence microscopy. Accurate PSF models account for the vector nature of light and the phase and amplitude modifications.…
Fluorescence Correlation Spectroscopy (FCS) yields measurement parameters (number of molecules, diffusion time) that characterize the concentration and kinetics of fluorescent molecules within a supposedly known observation volume. Absolute…
Fluorescent molecules are versatile nanoscale emitters that enable detailed observations of biophysical processes with nanoscale resolution. Because they are well-approximated as electric dipoles, imaging systems can be designed to…
Measuring subdiffraction separations between single fluorescent particles is important for biological, nano-, and medical-technology studies. Major challenges include (i) measuring changing molecular separations with high temporal…
Single molecule applications, saturated pattern excitation microscopy, or stimulated emission depletion (STED) microscopy demand for bright and highly stable fluorescent dyes1,2. Despite of intensive research the choice of fluorphores is…
Microstructured optical fibers (MOFs) are one of the most exciting recent developments in fiber optics. A MOF usually consists of a hexagonal arrangement of air holes running down the length of a silica fiber surrounding a central core of…
Fluorescence microscopy is widely used in biological imaging, however scattering from tissues strongly limits its applicability to a shallow depth. In this work we adapt a methodology inspired from stellar speckle interferometry, and…
Single-molecule approaches to biology offer a powerful new vision to elucidate the mechanisms that underpin the functioning of living cells. However, conventional optical single molecule spectroscopy techniques such as F\"orster…
The coupling of nanostructures with emitters opens ways for the realization of man-made subwavelength light emitting elements. In this article, we investigate the modification of fluorescence when an emitter is placed close to a…
Various techniques have been developed to measure the 2D and 3D positions and 2D and 3D orientations of fluorescent molecules with improved precision over standard epifluorescence microscopes. Due to the challenging signal-to-background…
Fluorescence Correlation Spectroscopy (FCS) is widely used to detect and quantify diffusion processes at the molecular level. The molecules of which diffusion is studied are marked with fluorescent dyes. It is commonly maintained that this…