Expression analysis of plvap in mouse heart development, homeostasis and injury.

Plasmalemma vesicle associated protein (PLVAP) is commonly considered to be specifically expressed in endothelial cells in which it localized to diaphragms of caveolae, fenestrae, and transendothelial channels. PLVAP is reported to be an important regulator of heart development and a novel target to promote cardiac repair in the ischemic heart. However, the dynamics of plvap expression in heart development, homeostasis and pathology have not been comprehensively described. In this study, we analyzed the temporal and spatial expression of plvap in mouse heart under different conditions. We found that, during embryonic and neonatal stages, PLVAP was detected in endocardial endothelial cells, epicardial mesothelial cells, and a small amount of coronary vascular endothelial cells. In adult heart, PLVAP was also identified in endocardial cells and a few coronary vascular endothelial cells. However, epicardial expression of PLVAP was lost during postnatal heart development and cannot be detected in mouse heart by immunostaining since 3-week-old. We also analyzed the expression of plvap in a model of cardiac hypertrophy and failure induced by transverse aortic constriction surgery, and identified expression of PLVAP in endocardial cells and coronary vascular endothelial cells in the injured heart. This study provides new evidence to better understand the role of plvap in mouse heart development and injury.

Advancing full-field metrology: rapid 3D imaging with geometric phase ferroelectric liquid crystal technology in full-field optical coherence microscopy.

Optical coherence microscopy (OCM) is a variant of OCT in which a high-numerical aperture lens is used. Full-field OCM (FF-OCM) is an emerging non-invasive, label-free, interferometric technique for imaging of surface structures or semi-transparent biomedical subjects with micron-scale resolutions. Different approaches to three dimensional full-field optical metrology are reviewed. The usual method for the phase-shifting technique in FF-OCM involves mechanically moving a mirror to change the optical path difference for obtaining en-face OCM images. However, with the use of a broadband source in FF-OCM, the phase shifts of different spectral components are not the same, resulting in the ambiguities in 3D image reconstruction. In this study, we demonstrate, by imaging tissues and cells, a unique geometric phase-shifter based on ferroelectric liquid crystal technology, to realize achromatic phase-shifting for rapid three-dimensional imaging in a FF-OCM system.

Genetic lineage tracing identifies cardiac mesenchymal-to-adipose transition in an arrhythmogenic cardiomyopathy model.

Arrhythmogenic cardiomyopathy (ACM) is one of the most common inherited cardiomyopathies, characterized by progressive fibrofatty replacement in the myocardium. However, the cellular origin of cardiac adipocytes in ACM remains largely unknown. Unraveling the cellular source of cardiac adipocytes in ACM would elucidate the underlying pathological process and provide a potential target for therapy. Herein, we generated an ACM mouse model by inactivating desmosomal gene desmoplakin in cardiomyocytes; and examined the adipogenic fates of several cell types in the disease model. The results showed that SOX9+, PDGFRa+, and PDGFRb+ mesenchymal cells, but not cardiomyocytes or smooth muscle cells, contribute to the intramyocardial adipocytes in the ACM model. Mechanistically, Bmp4 was highly expressed in the ACM mouse heart and functionally promoted cardiac mesenchymal-to-adipose transition in vitro.

Generation and characterization of a Myh6-driven Cre knockin mouse line.

Gene deletion by the Cre-Loxp system has facilitated functional studies of many critical genes in mice, offering important insights and allowing deeper understanding on the mechanisms underlying organ development and diseases, such as heart development and diseases. In this study, we generated a Myh6-Cre knockin mouse model by inserting the IRES-Cre-wpre-polyA cassette between the translational stop codon and the 3' untranslated region of the endogenous Myh6 gene. By crossing knockin mice with the Rosa26 reporter lines, we found that Myh6-Cre targeted cardiomyocytes at the embryonic and postnatal stages. In addition, we were able to inactivate the desmosome gene Desmoplakin (Dsp) by breeding Myh6-Cre mice with a conditional Dspflox knockout mouse line, which resulted in embryonic lethality during the mid-term pregnancy. These results suggest that the new Myh6-Cre mouse line can serve as a robust tool to dissect the distinct roles of genes involved in heart development and function.

No Evidence for Erythro-Myeloid Progenitor-Derived Vascular Endothelial Cells in Multiple Organs.

RATIONALE: Endothelial cells are thought to emerge de novo from the mesoderm to form the entire circulatory system. Recently, erythro-myeloid progenitors (EMPs) have been proposed to be another remarkable developmental origin for blood vessels in multiple organs, including the hindbrain, liver, lung, and heart, as demonstrated by lineage tracing studies using different genetic tools. These observations challenge the current consensus that intraembryonic vessels are thought to expand solely by the proliferation of preexisting endothelial cells. Resolution of this controversy over the developmental origin of endothelial cells is crucial for developing future therapeutics for vessel-dependent organ repair and regeneration. OBJECTIVE: To examine the contribution of EMPs to intraembryonic endothelial cells. METHODS AND RESULTS: We first used a transgenic mouse expressing a tamoxifen-inducible Mer-iCre fusion protein driven by the Csf1r (colony stimulating factor 1 receptor) promoter. Genetic lineage tracing based on Csf1r-Mer-iCre-Mer showed no contribution of EMPs to brain endothelial cells identified by several markers. We also generated a knock-in mouse line by inserting an internal ribosome entry site-iCre cassette into the 3' untranslated region of Csf1r gene to further investigate the cellular fates of EMPs. Similarly, we did not find any Csf1r-ires-iCre traced endothelial cells in brain, liver, lung, or heart in development either. Additionally, we found that Kit (KIT proto-oncogene receptor tyrosine kinase) was expressed not only in EMPs but also in embryonic hindbrain endothelial cells. Therefore, Kit promoter-driven recombinase, such as Kit-CreER, is a flawed tool for lineage tracing when examining the contribution of EMPs to hindbrain endothelial cells. We also traced CD45 (protein tyrosine phosphatase receptor type C; Ptprc)+ circulating EMPs and did not find any CD45 lineage-derived endothelial cells during development. CONCLUSIONS: Our study suggested that EMPs are not the origin of intraembryonic endothelial cells.

Active sorting of orbital angular momentum states of light with a cascaded tunable resonator.

The orbital angular momentum (OAM) of light has been shown to be useful in diverse fields ranging from astronomy and optical trapping to optical communications and data storage. However, one of the primary impediments preventing such applications from widespread adoption is the lack of a straightforward and dynamic method to sort incident OAM states without altering the states. Here, we report a technique that can dynamically filter individual OAM states and preserve the incident OAM states for subsequent processing. Although the working principle of this technique is based on resonance, the device operation is not limited to a particular wavelength. OAM states with different wavelengths can resonate in the resonator without any additional modulation other than changing the length of the cavity. Consequently, we are able to demonstrate a reconfigurable OAM sorter that is constructed by cascading such optical resonators. This approach does not require specially designed components and is readily amenable to integration into potential applications.

Dual lineage tracing identifies intermediate mesenchymal stage for endocardial contribution to fibroblasts, coronary mural cells, and adipocytes.

Early embryonic endocardium undergoes endothelial-to-mesenchymal transition to form cardiac cushion mesenchymal cells (MCs). Embryonic endocardium also gives rise to fibroblasts, intramyocardial adipocytes, and coronary mural cells, including smooth muscle cells and pericytes, in development. Whether endocardial cells directly differentiate into fibroblasts, coronary mural cells, and adipocytes or indirectly via an intermediate stage of endocardial-derived cushion MCs remains unknown. In addition to endocardium, epicardium and neural crest also contribute to cardiac cushion MCs. Given the developmental heterogeneity of cushion MCs and the lack of specific markers for endocardial-derived cushion MCs, conventional genetic lineage tracing utilizing Cre recombinase driven by one specific regulatory element is not sufficient to examine the fates of endocardial-derived cushion MCs. Intersectional genetic targeting approaches, which combine regulatory elements from two or more genes, have been employed to increase the specificity of cell targeting. Here, we developed a dual-recombinase intersectional targeting approach using Nfatc1-Dre, Sox9-CreER, and Cre/Dre double-dependent reporter Ai66 to specifically label endocardial-derived cushion MCs. Taking advantage of intersectional lineage tracing, we found that a subset of cardiac cells including fibroblasts, coronary mural cells, and intramyocardial adipocytes in adult hearts were derived from endocardial-derived cushion MCs. Our study suggests that embryonic endocardium contributes to cushion MCs first, and then endocardial-derived cushion MCs migrate into myocardium and differentiate into fibroblasts, coronary mural cells, and adipocytes in development. Understanding developmental origins of cardiac cell lineages will provide us more insights into cardiac development, regeneration, and diseases.

Toward broadband, dynamic structuring of a complex plasmonic field.

The ability to tailor a coherent surface plasmon polariton (SPP) field is an important step toward many new opportunities for a broad range of nanophotonic applications. Previously, both scanning a converging SPP spot and designing SPP profiles using an ensemble of spots have been demonstrated. SPPs, however, are normally excited by intense, coherent light sources, that is, lasers. Hence, interference between adjacent spots is inevitable and will affect the overall SPP field distributions. We report a reconfigurable and wavelength-independent platform for generating a tailored two-dimensional (2D) SPP field distribution by considering the coherent field as a whole rather than as individual spots. With this new approach, the inherent constraints in a 2D coherent field distribution are revealed. Our design approach works not only for SPP waves but also for other 2D wave systems such as surface acoustic waves.

Continuously Tunable, Polarization Controlled, Colour Palette Produced from Nanoscale Plasmonic Pixels.

Colour filters based on nano-apertures in thin metallic films have been widely studied due to their extraordinary optical transmission and small size. These properties make them prime candidates for use in high-resolution colour displays and high accuracy bio-sensors. The inclusion of polarization sensitive plasmonic features in such devices allow additional control over the electromagnetic field distribution, critical for investigations of polarization induced phenomena. Here we demonstrate that cross-shaped nano-apertures can be used for polarization controlled color tuning in the visible range and apply fundamental theoretical models to interpret key features of the transmitted spectrum. Full color transmission was achieved by fine-tuning the periodicity of the apertures, whilst keeping the geometry of individual apertures constant. We demonstrate this effect for both transverse electric and magnetic fields. Furthermore we have been able to demonstrate the same polarization sensitivity even for nano-size, sub-wavelength sets of arrays, which is paramount for ultra-high resolution compact colour displays.

On-chip photonic Fourier transform with surface plasmon polaritons.

The Fourier transform (FT), a cornerstone of optical processing, enables rapid evaluation of fundamental mathematical operations, such as derivatives and integrals. Conventionally, a converging lens performs an optical FT in free space when light passes through it. The speed of the transformation is limited by the thickness and the focal length of the lens. By using the wave nature of surface plasmon polaritons (SPPs), here we demonstrate that the FT can be implemented in a planar configuration with a minimal propagation distance of around 10 µm, resulting in an increase of speed by four to five orders of magnitude. The photonic FT was tested by synthesizing intricate SPP waves with their Fourier components. The reduced dimensionality in the minuscule device allows the future development of an ultrafast on-chip photonic information processing platform for large-scale optical computing.

Broadband chirality-coded meta-aperture for photon-spin resolving.

The behaviour of light transmitted through an individual subwavelength aperture becomes counterintuitive in the presence of surrounding 'decoration', a phenomenon known as the extraordinary optical transmission. Despite being polarization-sensitive, such an individual nano-aperture, however, often cannot differentiate between the two distinct spin-states of photons because of the loss of photon information on light-aperture interaction. This creates a 'blind-spot' for the aperture with respect to the helicity of chiral light. Here we report the development of a subwavelength aperture embedded with metasurfaces dubbed a 'meta-aperture', which breaks this spin degeneracy. By exploiting the phase-shaping capabilities of metasurfaces, we are able to create specific meta-apertures in which the pair of circularly polarized light spin-states produces opposite transmission spectra over a broad spectral range. The concept incorporating metasurfaces with nano-apertures provides a venue for exploring new physics on spin-aperture interaction and potentially has a broad range of applications in spin-optoelectronics and chiral sensing.

Mode-matching metasurfaces: coherent reconstruction and multiplexing of surface waves.

Metasurfaces are promising two-dimensional metamaterials that are engineered to provide unique properties or functionalities absent in naturally occurring homogeneous surfaces. Here, we report a type of metasurface for tailored reconstruction of surface plasmon waves from light. The design is based on an array of slit antennas arranged in a way that it matches the complex field distribution of the desired surface plasmon wave. The approach is generic so that one can readily create more intricate designs that selectively generate different surface plasmon waves through simple variation of the wavelength or the polarization state of incident light. The ultra-thin metasurface demonstrated in this paper provides a versatile interface between the conventional free-space optics and a two-dimensional platform such as surface plasmonics.

Two-dimensional complex source point solutions: application to propagationally invariant beams, optical fiber modes, planar waveguides, and plasmonic devices.

Highly convergent beam modes in two dimensions are considered based on rigorous solutions of the scalar wave (Helmholtz) equation, using the complex source point formalism. The modes are applicable to planar waveguide or surface plasmonic structures and nearly concentric microcavity resonator modes in two dimensions. A novel solution is that of a vortex beam, where the direction of propagation is in the plane of the vortex. The modes also can be used as a basis for the cross section of propagationally invariant beams in three dimensions and bow-tie-shaped optical fiber modes.

Temporal reshaping of two-dimensional pulses.

An analytic study of complete cylindrical focusing of pulses in two dimensions is presented, and compared with the analogous three-dimensional case of focusing over a complete sphere. Such behavior is relevant for understanding the limiting performance of ultrafast, planar photonic and plasmonic devices. A particular spectral distribution is assumed that contains finite energy. Separate ingoing and outgoing pulsed waves are considered, along with the combination that would be generated in free space by an ingoing wave. It is shown that for the two dimensional case, in order to produce a temporally symmetrical pulse at the focus, an asymmetric pulse must be launched. A symmetrical outgoing pulse is generated from a source with asymmetric time behavior, or an anti-symmetric input pulse. These results are very different from the corresponding three-dimensional case, and imply fundamental limitations on the performance of ultrafast, tightly focused, two-dimensional devices.

Calculation of the volumetric diffracted field with a three-dimensional convolution: the three-dimensional angular spectrum method.

The first Rayleigh-Sommerfeld diffraction formula is treated in an exact form as a three-dimensional (3D) convolution in the spatial domain. Therefore, a 3D Fourier transform can be employed to convert the 3D diffracted electromagnetic field to the reciprocal space without approximations, which we call the 3D angular spectrum (3D-AS) method. It is also demonstrated that if evanescent waves are neglected, the 3D-AS method can be readily implemented numerically, with the results in good agreement with theoretical predictions.

Reconstruction in interferometric synthetic aperture microscopy: comparison with optical coherence tomography and digital holographic microscopy.

It is shown that the spatial frequencies recorded in interferometric synthetic aperture microscopy do not correspond to exact backscattering [as they do in unistatic synthetic aperture radar (SAR)] and that the reconstruction process based on SAR is therefore based on an approximation. The spatial frequency response is developed based on the three-dimensional coherent transfer function approach and compared with that in optical coherence tomography and digital holographic microscopy.

Realistic 3D coherent transfer function inverse filtering of complex fields.

We present a novel technique for three-dimensional (3D) image processing of complex fields. It consists in inverting the coherent image formation by filtering the complex spectrum with a realistic 3D coherent transfer function (CTF) of a high-NA digital holographic microscope. By combining scattering theory and signal processing, the method is demonstrated to yield the reconstruction of a scattering object field. Experimental reconstructions in phase and amplitude are presented under non-design imaging conditions. The suggested technique is best suited for an implementation in high-resolution diffraction tomography based on sample or illumination rotation.

Quantitative phase restoration by direct inversion using the optical transfer function.

Quantitative phase recovery of phase objects is achieved by a direct inversion using the defocused weak object transfer function. The presented method is noniterative and is based on partially coherent principles. It also takes into account the optical properties of the system and gives the phase of the object directly. The proposed method is especially suitable for application to weak phase objects, such as live and unstained biological samples but, surprisingly, has also been shown to work with comparatively strong phase objects.