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Light microscopes are the most widely used devices in life and material sciences that allow the study of the interaction of light with matter at a resolution better than that of the naked eye. Conventional microscopes translate the spatial differences in the intensity of the reflected or transmitted light from an object to pixel brightness differences in the digital image. However, a phase microscope converts the spatial differences in the phase of the light from or through an object to differences in pixel brightness. Interference microscopy, a phase-based approach, has found application in various disciplines. While interferometry has brought nanometric axial resolution, the lateral resolution in quantitative phase microscopy (QPM) has still remained limited by diffraction, similar to other traditional microscopy systems. Enhancing the resolution has been the subject of intense investigation since the invention of the microscope in the 17th century. During the past decade, microsphere-assisted microscopy (MAM) has emerged as a simple and effective approach to enhance the resolution in light microscopy. MAM can be integrated with QPM for 3D label-free imaging with enhanced resolution. Here, we review the integration of microspheres with coherence scanning interference and digital holographic microscopies, discussing the associated open questions, challenges, and opportunities.

Shaped optical fiber tips have recently attracted a lot of interest for photonic jet light focusing due to their easy manipulation to scan a sample. However, lensed optical fibers are not new. This study analyzes how fiber tip parameters can be used to control focusing properties. Our study shows that the configurations to generate a photonic jet (PJ) can clearly be distinguished from more classical-lensed fibers focusing. PJ is a highly concentrated, propagative light beam, with a full width at half maximum (FWHM) that can be lower than the diffraction limit. According to the simulations, the PJs are obtained when light is coupled in the guide fundamental mode and when the base diameter of the microlens is close to the core diameter. For single mode fibers or fibers with a low number of modes, long tips with a relatively sharp shape achieve PJ with smaller widths. On the contrary, when the base diameter of the microlens is larger than the fiber core, the focus point tends to move away from the external surface of the fiber and has a larger width. In other words, the optical system (fiber/microlens) behaves in this case like a classical-lensed fiber with a larger focus spot size. The results of this study can be used as guidelines for the tailored fabrication of shaped optical fiber tips according to the targeted application.

Abstract We present quantitative three dimensional images of grooves on a writable Blu-ray Disc based on a single objective Mirau type interferometric microscope, enhanced with a microsphere which is considered as a photonic nanojet source. Along the optical axis the resolution of this microsphere assisted interferometry system is a few nanometers while the lateral resolution is around 112 nm. To understand the physical phenomena involved in this kind of imaging we have modelled the interaction between the photonic jet and the complex disc surface. Agreement between simulation and experimental results is demonstrated. We underline that although the ability of the microsphere to generate a photonic nanojet does not alone explain the resolution of the interferometer, the nanojet can be used to try to understand the imaging process. To partly explain the lateral super-resolution, the potential role of coherence is illustrated. The presented modality may have a large impact on many fields from bio-medicine to nanotechnology.

Through rigorous electromagnetic simulations, the natural coupling of high-spatial-frequency evanescent waves from the near field to the far field by dielectric microspheres is studied in air. The generation of whispering gallery modes inside the microspheres is shown independently of any resonance. In addition, the conversion mechanism of these evanescent waves into propagating waves is analysed. This latter point leads to key information that allows a better physical understanding of the super-resolution phenomenon in microsphere-assisted microscopy where sub-diffraction-limit revolving power is achieved.

The role of genetics in the development of osteoarthritis is well established but the molecular bases are not fully understood. Here, we describe a family carrying a germline mutation in COMP (Cartilage Oligomeric Matrix Protein) associated with three distinct phenotypes. The index case was enrolled for a familial form of idiopathic early-onset osteoarthritis. By screening potential causal genes for osteoarthritis, we identified a heterozygous missense mutation of COMP (c.1358C>T, p.Asn453Ser), absent from genome databases, located on a highly conserved residue and predicted to be deleterious. Molecular dynamics simulation suggests that the mutation destabilizes the overall COMP protein structure and consequently the calcium releases from neighboring calcium binding sites. This mutation was once reported in the literature as causal for severe multiple epiphyseal dysplasia (MED). However, no sign of dysplasia was present in the index case. The mutation was also identified in one of her brothers diagnosed with MED and secondary osteoarthritis, and in her sister affected by an atypical syndrome including peripheral inflammatory arthritis of unknown cause, without osteoarthritis nor dysplasia. This article suggests that this mutation of COMP is not only causal for idiopathic early-onset osteoarthritis or severe MED, but can also be associated to a broad phenotypic variability with always joint alterations.

Abstract 3-Deazaneplanocin A (DZNep) is an inhibitor of S-Adenosyl-L-Homocysteine Hydrolase (SAHH) known to inhibit EZH2, a histone methylase upregulated during osteoarthritis. In this study, we assessed its effects in human articular chondrocytes. Anti-inflammatory effects were assessed by Nitric Oxide (NO), Prostaglandin E2 (PGE2) and Metalloprotease (MMP) release in IL-1β-stimulated chondrocytes. MAPK and NFκB activation was analyzed by western blotting. Differentially expressed genes (DEG) regulated by DZNep were identified by whole-transcriptome microarray. DZNep inhibited SAHH activity and was not toxic. It counteracted NO, PGE2 and MMP release, and reduced MAPK activation induced by IL-1β. By whole-transcriptome analysis, we identified that DNZep counteracts the effect of IL-1β on the expression of 81 protein-coding genes, including CITED2, an MMP inhibitor. These genes are organized in a protein-protein network centred on EGR1, which is known to functionally interact with EZH2. Gene ontologies enrichment analysis confirmed that DZNep counteracts IL-1β-induced expression of genes involved in cartilage matrix breakdown (MMPs and ADAMTS). In addition, DZNep up-regulated cartilage specific genes, such as COL2A1 and SOX9, suggesting a chondroprotective effect of DZNep. DZNep exhibits anti-inflammatory effects, and regulates genes implicated in chondroprotective response in human articular chondrocytes, suggesting that inhibitors of S-adenosylmethionine-dependent methyltransferases could be effective treatments for OA.

Abstract Mesenchymal stem cells (MSCs) hold promise for cartilage engineering. Here, we aimed to determine the best culture conditions to induce chondrogenesis of MSCs isolated from bone marrow (BM) of aged osteoarthritis (OA) patients. We showed that these BM-MSCs proliferate slowly, are not uniformly positive for stem cell markers, and maintain their multilineage potential throughout multiple passages. The chondrogenic lineage of BM-MSCs was induced in collagen scaffolds, under normoxia or hypoxia, by BMP-2 and/or TGF-β1. The best chondrogenic induction, with the least hypertrophic induction, was obtained with the combination of BMP-2 and TGF-β1 under hypoxia. Differentiated BM-MSCs were then transfected with siRNAs targeting two markers overexpressed in OA chondrocytes, type I collagen and/or HtrA1 protease. siRNAs significantly decreased mRNA and protein levels of type I collagen and HtrA1, resulting in a more typical chondrocyte phenotype, but with frequent calcification of the subcutaneously implanted constructs in a nude mouse model. Our 3D culture model with BMP-2/TGF-β1 and COL1A1/HtrA1 siRNAs was not effective in producing a cartilage-like matrix in vivo. Further optimization is needed to stabilize the chondrocyte phenotype of differentiated BM-MSCs. Nevertheless, this study offers the opportunity to develop a combinatory cellular therapy strategy for cartilage tissue engineering.

The direct simultaneous acquisition of coherent imaging and strain information is of particular importance in the biomechanical characterization of biological tissue. This type of simultaneous information acquisition can be accomplished using a coupled photorefractive holography and shearography system for imaging and strain measurements, respectively. Optical scattering in a conventional speckle shearing interferometer rapidly reduces the contrast of the shearing fringes, thereby limiting the use of such interferometers with opaque surfaces. By coupling photorefractive holography with speckle shearing interferometry, properties of the photorefractive effect (spatial high-pass filtering and temporal low-pass filtering) combine to restore the shearing fringe contrast and enable strain imaging in diffusing media. This effect is demonstrated using synthetic scattering phantoms built from suspensions of silica spheres in water. [ABSTRACT FROM AUTHOR]

A photonic nanojet is a highly localized electromagnetic propagative beam in the shadow side of a dielectric micro-particle. This concept was introduced in a famous paper by Chen et al. in 2004. Since then, this paper has inspired numerous original developments all over the world. Several years later, we point out that whispering gallery mode (WGM) resonances, not always brought out by FDTD simulations, can coexist in addition to the photonic jet. Such WGM resonances drastically change the distribution of the electric field. Through subsequent simulations, we show how these WGM resonances are highly sensitive to parameter changes. Particle diameter change smaller than 1 nm is enough to make them disappear. This may explain why sometimes they are not observed by FDTD simulations despite high resolution spatial mesh.