Miroslav Stibůrek | Searcharxiv

arXiv2025

Funnelling super-resolution STED microscopy through multimode fibres

Holographic multimode fibre endoscopes have recently shown their ability to unveil and monitor deep brain structures with sub-micrometre resolution, establishing themselves as a minimally-invasive technology with promising applications in neurobiology. In this approach, holographic control of the input light field entering the multimode fibres is achieved by means of wavefront shaping, usually treating the fibre as a complex medium. In contrast to other unpredictable and highly scattering complex media, multimode fibres feature symmetries and strong correlations between their input and output fields. Both step-index and graded-index multimode fibres offer a specific set of such correlations which, when appropriately leveraged, enable generating high-quality focused pulses with minimal intermodal dispersion. With this, we funnelled pulsed super-resolution STED microscopy with time-gated detection through a custom multimode fibre probe, combining the correlations of both multimode fibre types. We demonstrate resolution improvements over 3-times beyond the diffraction limit and showcase its applicability in bioimaging. This work provides not only a solution for delivering short pulses through step-index multimode fibre segments but also marks a step towards bringing advanced super-resolution imaging techniques with virtually no depth limitations.

physics.optics↗

arXiv2025

Hair-thin confocal fluorescence endo-microscopy for deep-brain in-vivo imaging

Confocal and multi-photon microscopy are widely used for in-vivo fluorescence imaging of biological tissues such as the brain, offering non-invasive access up to ~1 mm depth without major loss in performance. A recently-developed alternative is holographic endoscopy, which exploits controlled light transport through hair-thin optical fibres. With minimal invasiveness, it provides observations at comparable spatial resolution, while extending its applicability to unprecedented depths. It has been used to resolve details of sub-cellular structural connectivity, record neuronal signalling, and monitor blood flow from the deepest locations of the living brain. Yet, its use, particularly in densely labelled brain regions, has so far been constrained by significant contrast loss, primarily due to the absence of a practical mechanism for rejecting out-of-focus fluorescence light -- a capability inherently provided by confocal and multi-photon microscopy. Exploring opportunities in the structure of light modes of different MMF types we identify the possibility of achieving an analogue to confocal fluorescence microscopy through MMF-based endoscopes. Using a novel composite fibre probe that combines graded-index and step-index MMFs, we enable spatially resolved signal collection and selective rejection of out-of-focus light. This confocal filtering significantly enhances image contrast and resolution by suppressing background and off-plane signals. We demonstrate improved imaging performance on fine structural connectivity and intracellular calcium signalling in living mouse brain.

physics.optics↗