Abstract: Label-free microscopy exploits light scattering to obtain a three-dimensional image of biological tissues. However, light propagation is affected by aberrations and multiple scattering, which drastically degrade the image quality and limit the penetration depth. Multi-conjugate adaptive optics and time-gated matrix approaches have been developed to compensate for aberrations but the associated frame rate is extremely limited for three-dimensional imaging. Here we develop a multi-spectral matrix approach to solve these fundamental problems. On the basis of a sparse illumination scheme and an interferometric measurement of the reflected wave field at multiple wavelengths, the focusing process can be optimized in post-processing for any voxel by addressing independently each frequency component of the reflection matrix. A proof-of-concept experiment shows a three-dimensional image of an opaque human cornea over a 0.1 mm3 field of view at a 290 nm resolution and a 1 Hz frame rate. This work paves the way towards a fully digital microscope allowing real-time, in vivo, quantitative and deep inspection of tissues.

Abstract: In an ideal perfectly straight multimode fiber with a circular core, the symmetry ensures that rotating the input wave front leads to a corresponding rotation of the output wave front. This invariant property, known as the rotational memory effect (RME), remains independent of the typically unknown output profile. The RME thus offers significant potential for imaging and telecommunication applications. However, in real-life fibers, this effect is degraded by intrinsic imperfections and external perturbations, and is challenging to observe because of its acute sensitivity to misalignments and aberrations in the optical setup. Building on a previously established method for precisely estimating fiber transmission properties, we demonstrate an accurate extraction of RME properties. Additionally, we introduce a comprehensive theoretical framework for both qualitative and quantitative analysis, which specifically links the angular-dependent correlation of the RME to the core deformation’s geometrical properties and the fiber’s mode characteristics. Our theoretical predictions align well with experimental data and simulations for various amounts of fiber distorsion. Finally, we demonstrate the ability to engineer wave fronts with significantly enhanced correlation across all rotation angles. This work enables accurate characterization of distributed disorder from the fabrication process and facilitates calibration-free imaging in multimode fibers.

Abstract: Purpose: Putative microglia were recently detected using adaptive optics ophthalmoscopy in healthy eyes. Here we evaluate the use of nonconfocal adaptive optics scanning light ophthalmoscopy (AOSLO) for quantifying the morphology and motility of presumed microglia and other immune cells in eyes with retinal inflammation from uveitis and healthy eyes. Design: Observational exploratory study. Participants: Twelve participants were imaged, including 8 healthy participants and 4 posterior uveitis patients recruited from the clinic of 1 of the authors (M.H.E.). Methods: The Pittsburgh AOSLO imaging system was used with a custom-designed 7-fiber optical fiber bundle for simultaneous confocal and nonconfocal multioffset detection. The inner retina was imaged at several locations at multiple timepoints in healthy participants and uveitis patients to generate time-lapse images. Main Outcome Measures: Microglia and macrophages were manually segmented from nonconfocal AOSLO images, and their morphological characteristics quantified (including soma size, diameter, and circularity). Cell soma motion was quantified across time for periods of up to 30 minutes and their speeds were calculated by measuring their displacement over time. Results: A spectrum of cell morphologies was detected in healthy eyes from circular amoeboid cells to elongated cells with visible processes, resembling activated and ramified microglia, respectively. Average soma diameter was 16.1 ± 0.9 μm. Cell movement was slow in healthy eyes (0.02 μm/sec on average), but macrophage-like cells moved rapidly in some uveitis patients (up to 3 μm/sec). In an eye with infectious uveitis, many macrophage-like cells were detected; during treatment their quantity and motility decreased as vision improved. Conclusions: In vivo adaptive optics ophthalmoscopy offers promise as a potentially powerful tool for detecting and monitoring inflammation and response to treatment at a cellular level in the living eye. Financial Disclosure(s): Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.