KEYWORDS: Tissues, Objectives, Image resolution, Light sources and illumination, Microscopy, Visualization, Lenses, In vivo imaging, Point spread functions, Optical microsystems
We have developed a compact scattering-based light sheet microscopy (sLSM) probe capable of imaging unstained tissues with cellular resolution. In the compact sLSM probe, a custom miniature objective lens was developed to achieve a high lateral resolution, large field of view (FOV), and small field curvature. The measured resolution of the custom objective was 1.65 to 1.97 μm across the FOV of ±1.0 mm. The compact probe had dimensions of 4 cm in width and height and 10 cm in length. The compact sLSM probe achieved an axial resolution better than 5.6 μm over a depth range of 206.2 μm and a lateral resolution of 1.9 μm. Preliminary results showed that the compact sLSM probe could visualize cellular details of fixed human anal epithelial tissues in a similar manner to a bench light sheet microscopy device using off-the-shelf objective lenses.
A general approach for modeling the temperature dependence of optical absorptions in rare earth-doped crystals is presented and applied to transitions in Tm3+:YAG. The model allows for the generation of calculated absorption spectra at any temperature. There is close agreement between the model and measurements conducted of optical absorption in Tm3+:YAG for the 3F3 and 3H4 manifolds between 50 and 300 K. A model of the temperature variation of optical absorption features is highly useful in the design of high-powered solid-state laser systems. In addition, it is shown that absorption into Tm3+:YAG can be used as an optical thermometer at cryogenic temperatures. Absorption data as a function of wavelength and temperature was obtained for a sample of 1% Tm3+:YAG, which was mounted inside a cryostat with optical access. Broadband light from a tungsten-filament lamp passed through the crystal and entered a 0.25 m monochrometer and was detected with a photomultiplier tube. Transmitted light was analyzed for two spectral ranges: 650 - 725 nm which includes absorption into the 3F3 manifold, and 750 – 825 nm which includes absorption into the 3H4 manifold. The data was compared to a model for absorption cross sections that includes the temperature dependence of homogeneous broadening mechanisms and thermalization within the ground state Stark levels. This approach to modeling the temperature dependence of optical absorption should have general applicability for other rare earth-doped crystals.
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