Purpose

High resolution visualization of the ocular surface and intact eyeball is critical and essential for our understanding and treatment of eye diseases. This study is to achieve this goal using advanced tissue clearing and three-dimensional (3D) imaging technologies.

Methods

Wild type and fluorescently labeled transgenic mice of Prox-1-GFP (green fluorescent protein) or Thy1-YFP (yellow fluorescent protein) were used in the study. Eyeballs were harvested from normal or a disease model of corneal inflammation. Samples were infused with hydrogel monomers and heated for polymerization. Lipids were removed by electrophoresis. The transparent tissue-hydrogel hybrids of the anterior segments or intact eyeballs with immunolabeling or endogenous fluorescence were imaged by an advanced light sheet fluorescent microscope. High resolution 3D images and videos were captured for a wide array of structures and cell types.

Results

Optical transparency was achieved from intact eyeballs of both normal and diseased conditions. A variety of important structures and cell types, such as blood and lymphatic vessels, Schlemm's canal, nerves and endothelial cells, were detected with their natural morphology, location and organizational network.

Conclusions

This study provides the first comprehensive and 3D high resolution imaging of the intact eyeball using tissue clearing and advanced light sheet microscopy. Given that the eye is the window of the body, we anticipate this advanced technology will facilitate diverse applications in biomedical research inside and outside the eye.

How does experiencing a rare event, like a car accident or a lottery win, influence decision-makers consecutive decisions? Studying these so-called recency effects holds a long tradition in research on experience-based decision-making. Previous work suggests opposite behavioral patterns after experiencing a positive rare event: People have been found to be more likely to either repeat their choice (positive recency) or to avoid it (negative recency). The effect is thought to persevere for multiple choices and decrease over time. In this study, we provide new insights into recency effects by analyzing people's repeated choices from an extensive database—consisting of 3 million choices by 8,000 participants across 12 different decision-from-experience paradigms collected from 139 studies. We provide a conceptual framework clarifying patterns of positive and negative recency, including how the direction and magnitude of impact change over time.

PURPOSE: Contact lens wear induces corneal parainflammation involving increased immune cell numbers after 24 hours (CD11c+, Lyz2+, γδ-T cells) and six days (Ly6G+ cells) wear. We investigated the time course of onset and resolution of these responses. METHODS: LysMcre or C57BL/6J mice were fitted with a contact lens (four to 48 hours). Contralateral eyes did not wear lenses. After lens removal, Lyz2+, MHC-II+ or Ly6G+ cells were examined by quantitative imaging. RT-qPCR determined cytokine gene expression. RESULTS: Lens wear for 24 hours increased corneal Lyz2+ cells versus contralateral eyes approximately two-fold. Corneas remained free of visible pathology. The Lyz2+ response was not observed after four or 12 hours wear, nor after 12 hours wear plus 12 hours no wear. Lens removal after 24 hours wear further increased Lyz2+ cells (∼48% after one day), which persisted for four days, returning to baseline by seven days. Lyz2+ cells in contralateral eyes remained at baseline. MHC-II+ cells showed a similar response but without increasing after lens removal. Lens wear for 48 hours showed reduced Lyz2+ cells versus 24 hours wear with one day discontinuation, correlating with reduced IL-1β and IL-18 gene expression. Lens wear for 24 hours did not induce Ly6G+ responses six days after removal. CONCLUSIONS: Lens-induced corneal parainflammation involving Lyz2+ cells requires 24 hours wear but persists after lens discontinuation, requiring seven days for reversal. Lens wear for 48 hours may suppress initial Lyz2+ cell and cytokine responses. The significance of parainflammation during and after lens wear remains to be determined.

Ultrafast, high-intensity light-matter interactions lead to optical-field-driven photocurrents with an attosecond-level temporal response. These photocurrents can be used to detect the carrier-envelope-phase (CEP) of short optical pulses, and enable optical-frequency, petahertz (PHz) electronics for high-speed information processing. Despite recent reports on optical-field-driven photocurrents in various nanoscale solid-state materials, little has been done in examining the large-scale electronic integration of these devices to improve their functionality and compactness. In this work, we demonstrate enhanced, on-chip CEP detection via optical-field-driven photocurrents in a monolithic array of electrically-connected plasmonic bow-tie nanoantennas that are contained within an area of hundreds of square microns. The technique is scalable and could potentially be used for shot-to-shot CEP tagging applications requiring orders-of-magnitude less pulse energy compared to alternative ionization-based techniques. Our results open avenues for compact time-domain, on-chip CEP detection, and inform the development of integrated circuits for PHz electronics as well as integrated platforms for attosecond and strong-field science.

Glycogen synthase kinase 3 (GSK3) plays a central role in diverse cellular processes. GSK3 has two mammalian isozymes, GSK3α and GSK3β, whose functions remain ill-defined because of a lack of inhibitors that can distinguish between the two highly homologous isozymes. Here, we show that GSK3α and GSK3β can be selectively inhibited in mouse embryonic stem cells (ESCs) using a chemical-genetic approach. Selective inhibition of GSK3β is sufficient to maintain mouse ESC self-renewal, whereas GSK3α inhibition promotes mouse ESC differentiation toward neural lineages. Genome-wide transcriptional analysis reveals that GSK3α and GSK3β have distinct sets of downstream targets. Furthermore, selective inhibition of individual GSK3 isozymes yields distinct phenotypes from gene deletion, highlighting the power of the chemical-genetic approach in dissecting kinase catalytic functions from the proteins scaffolding functions. Our study opens new avenues for defining GSK3 isozyme-specific functions in various cellular processes.

Despite high metabolic activity, the retina and optic nerve head lack traditional lymphatic drainage. We here identified an ocular glymphatic clearance route for fluid and wastes via the proximal optic nerve in rodents. β-amyloid (Aβ) was cleared from the retina and vitreous via a pathway dependent on glial water channel aquaporin-4 (AQP4) and driven by the ocular-cranial pressure difference. After traversing the lamina barrier, intra-axonal Aβ was cleared via the perivenous space and subsequently drained to lymphatic vessels. Light-induced pupil constriction enhanced efflux, whereas atropine or raising intracranial pressure blocked efflux. In two distinct murine models of glaucoma, Aβ leaked from the eye via defects in the lamina barrier instead of directional axonal efflux. The results suggest that, in rodents, the removal of fluid and metabolites from the intraocular space occurs through a glymphatic pathway that might be impaired in glaucoma.