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Three-Dimensional Segmentation and Quantitative Measurement of the Aqueous Outflow System of Intact Mouse Eyes Based on Spectral Two-Photon Microscopy Techniques
摘要： PURPOSE. To visualize and quantify the three-dimensional (3D) spatial relationships of the structures of the aqueous outflow system (AOS) within intact enucleated mouse eyes using spectral two-photon microscopy (TPM) techniques. METHODS. Spectral TPM, including two-photon autofluorescence (TPAF) and second-harmonic generation (SHG), were used to image the small structures of the AOS within the limbal region of freshly enucleated mouse eyes. Long infrared excitation wavelengths (930 nm) were used to reduce optical scattering and autofluorescent background. Image stacks were collected for 3D image rendering and structural segmentation. For anatomical reference, vascular perfusion with fluorescent-conjugated dextran (150 KDa) and trans-corneal perfusion with 0.1 lm fluorescent polystyrene beads were separately performed to identify the episcleral veins (EV) and the trabecular meshwork (TM) and Schlemm's canal (SC), respectively. RESULTS. Three-dimensional rendering and segmentation of spectral two-photon images revealed detailed structures of the AOS, including SC, collector channels (CC), and aqueous veins (AV). The collagen of the TM was detected proximal to SC. The long and short axes of the SC were 82.2 ± 22.2 μm and 6.7 ± 1.4 μm. The diameters of the CC averaged 25.6 ± 7.9 μm where they originated from the SC (ostia), enlarged to 34.1 ± 13.1 μm at the midpoint, and narrowed to 18.3 ± 4.8 μm at the junction of the AV. The diameter of the AV averaged 12.5 ± 3.4 μm. CONCLUSIONS. Spectral TPM can be used to reconstruct and measure the spatial relationships of both large and small AOS structures, which will allow for better understanding of distal aqueous outflow dynamics.
关键词： second-harmonic generation,aqueous outflow,nonlinear microscopy,glaucoma,two-photon fluorescence microscopy
更新于2025-11-21 11:08:12
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[IEEE 2019 Conference on Lasers and Electro-Optics Europe & European Quantum Electronics Conference (CLEO/Europe-EQEC) - Munich, Germany (2019.6.23-2019.6.27)] 2019 Conference on Lasers and Electro-Optics Europe & European Quantum Electronics Conference (CLEO/Europe-EQEC) - Nonlinear Microscopy of Lead Iodide Nanosheets
摘要： Lead iodide (PbI2) is a layered material with unique optical and electrical properties, including direct bandgap in the bulk and a layered crystalline structure, consisting of close-packed Pb atoms sandwiched between two layers of Iodine atoms. Compared to the widely-studied TMDCs, 2D PbI2 is a new type of halide semiconductors with a relatively larger visible bandgap (Eg ～ 2.4 eV) which endows its distinct optical properties. Despite being studied for decades and being used as a precursor for perovskite materials, the recently developed PbI2 nanosheets have shown a great promise for high-performance optoelectronic devices, such as ?exible photodetectors and nanolasers. Such 2D nanosheets also show a great potential for low-dimensional nonlinear optical devices, However, their nonlinear properties are still unexplored, while novel applications of PbI2 nanosheets require careful characterization of their crystalline structure, thickness and nonlinearity. Here we demonstrate the nonlinear microscopy of PbI2 nanosheets using the polarization and thickness dependence of the second harmonic generation (SHG) and third harmonic generation (THG) from solution-grown nanosheets. Our measurements allow to precisely determine their thickness and crystalline orientation with a non-invasive optical technique.
关键词： third harmonic generation,nonlinear microscopy,Lead iodide,second harmonic generation,nanosheets
更新于2025-09-11 14:15:04