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Martin Oheim

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Published work

4 published item(s)

preprint2026arXiv

Label-Free Microrefractometry of Interfacial Processes Using Fluorescent Smart Coverslips

Molecular dipoles near interfaces emit highly directional radiation due to near-field interactions, making surface-bound fluorophores sensitive probes of local physicochemical changes. We introduce smart coverslips, stably coated with uniform, brightly fluorescent nanobead films, that exploit refractive-index-dependent emission shifts for sensitive micro-refractometry in small volumes. Supercritical-angle fluorescence refractometry uses single back-focal-plane images to allow us real-time RI sensing and nanometric thin-film height measurements without the need for multi-angle or multi-wavelength acquisition. Our fast, label-free, and non-invasive approach allows measurements of thin-film properties and monitoring of interfacial dynamics on a standard inverted microscope and is broadly applicable to nanobiophotonics, chemical sensing, and in-situ materials analysis.

preprint2026arXiv

Universal Nano-Bead Emitter Inks for Programmable Nanometric Fluorescent Architectures

Fabricating brightly fluorescent layers with nanometric thickness and digitally controlled lateral structuration remains a challenge for next-generation photonic devices, optical calibration standards, and biocompatible interfaces. Here, we introduce Nano-Bead Emitters (NBEs), hydrogel nanoparticles covalently functionalized with fluorophores, as a universal, water-processable ink platform for fabricating programmable nanometric fluorescent architectures. By immobilizing fluorophores within a charged nanohydrogel scaffold, the platform entirely decouples film morphology from dye solubility. This molecule-independent strategy enables spectrally distinct, inherently water-insoluble dyes to be processed using a single, standardized aqueous ink formulation. Combined with laser-induced forward transfer (LIFT) printing, this additive approach yields highly uniform fluorescent layers (~7 nm thickness, sub-nanometric roughness). This structural invariance produces complex multicolor patterns sharing identical thickness and surface morphology across all spectral channels, a critical requirement for quantitative optical calibration. Furthermore, LIFT printing provides programmable, layer-by-layer control over fluorescence intensity via successive deposition cycles, yielding precisely tunable brightness without aggregation-caused quenching. This maskless technique enables rapid, high-fidelity printing of both monochromatic and multicolor patterns over macroscopic areas with absolute spatial resolution. Finally, these universally compatible NBE inks stably deposit onto diverse substrates (glass, polymers, semiconductors, metasurfaces), effectively bridging scalable manufacturing with high-performance integrated photonic systems.

preprint2020arXiv

Supercritical angle microscopy and spectroscopy

Fluorescence detection, either involving propagating or near-field emission, is widely being used in spectroscopy, sensing and microscopy. Total internal reflection fluorescence (TIRF) confines fluorescence excitation by an evanescent (near-) field and it is a popular contrast generator for surface-selective fluorescence assays. Its emission equivalent, supercritical angle fluorescence (SAF) is comparably less established although it achieves a similar optical sectioning as does TIRF. SAF emerges when a fluorescing molecule is located very close to an interface and its near-field emission couples to the higher-refractive index medium (n2 > n1) and becomes propagative. Then, most fluorescence is detectable on the side of the higher-index substrate and a large fraction of this fluorescence is emitted into angles forbidden by Snell's law. SAF as well as the undercritical angle fluorescence (UAF) (far-field emission) components can be collected with microscope objectives having a high-enough detection aperture NA > n2 and be separated in the back-focal plane (BFP) by Fourier filtering. The BFP image encodes information about the fluorophore radiation pattern, and it can be analysed to yield precise information about the refractive index in which the emitters are embedded, their nanometric distance from the interface and their orientation. A SAF microscope can retrieve this near-field information through wide-field optics in a spatially resolved manner, and this functionality can be added to any existing inverted microscope.

preprint2013arXiv

Combined evanescent-wave excitation and supercritical-angle fluorescence detection improves optical sectioning

Evanescent-wave microscopy achieves sub-diffraction axial sectioning by confining fluorescence excitation to a thin layer close to the cell/substrate interface. How thin this light sheet exactly is, however, is often unknown. Particularly in the popular objective-type total internal reflection fluorescence microscopy (TIRFM) configuration large deviations from the expected exponential intensity decay of the evanescent wave have been reported. Propagating, i.e., non-evanescent, excitation light diminishes the optical sectioning effect, reduces contrast and renders the quantification of TIRFM images uncertain. Here, we use a combination of azimuthal- and polar-angle beam scanning, dark-field scatter imaging, and atomic force microscopy to identify the sources of this unwanted background fluorescence excitation. We identify stray light originating from the microscope optics and the objective lens itself as the major sources of background, with minor contributions due to evanescent-wave scattering at the reflecting interface and at refractive-index boundaries in the sample. Apart from evanescence in excitation light, light emitted from a fluorophore can also show observable effects of evanescence. Only fluorophores located close to the coverslip can couple their near-field radiation into propagating waves detectable at supercritical angles. We show that selectively detecting this supercritical-angle fluorescence (SAF) through a high-numerical aperture objective effectively rejects fluorescence from deeper sample regions and improves optical sectioning. The microscopy scheme presented here merges the benefits of TIRF excitation and SAF detection and provides the conditions for quantitative wide-field imaging of fluorophore dynamics at or near the plasma membrane.