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Fig. 5 | BMC Biology

Fig. 5

From: Light sheet theta microscopy for rapid high-resolution imaging of large biological samples

Fig. 5

Rapid uniform high-resolution imaging of mouse central nervous system. a A CLARITY-cleared Thy1-eYFP transgenic mouse brain with attached spinal cord was imaged with LSTM microscopy using 10×/0.6NA/8mmWD detection objective (correction collar adjusted to 1.45 refractive index). A rolling shutter exposure of 0.5 ms and a full frame exposure of 20 ms were used. High-resolution 3D rendering was generated after 2 × 2-fold down-sampling. The bounding boxes are 11.8 mm × 27.6 mm × 5.2 mm for the whole sample and 5.1 mm × 3.1 mm × 3.5 mm for the subvolume (magenta). Images were acquired with 5-μm z-spacing using an effective light sheet thickness of ~ 5 μm. Lateral pixel sampling was 0.585 × 0.585 μm. A detailed volume rendering is shown in Additional file 8: Video 3 [29]. b A large thick coronal slice of a Thy1-eYFP transgenic mouse brain was imaged with LSTM using 488 nm excitation wavelength. A rolling shutter exposure window of 0.5 ms and a full frame exposure of 20 ms were used. The volume rendering was performed using 4 × 4 fold down-sampled data. The bounding box is 9.6 mm × 13.5 mm × 5.34 mm. Images were acquired with 5-μm z-spacing using an effective light sheet thickness of ~ 5 μm. Lateral pixel sampling was 0.585 × 0.585 μm. Additional file 9: Video 4 [29] shows volumetric rendering. c The same sample as shown in b was imaged with a high-NA 25×/1.0NA/8mmWD objective. A rolling shutter exposure window of 0.4 ms and a full frame exposure of 20 ms were used. The volume rendering was performed after 2 × 2-fold down-sampling. The bounding box is 6 mm × 9.6 mm × 0.5 mm. Images were acquired with 5-μm z-spacing using an effective light sheet thickness of ~ 3 μm. Lateral pixel sampling was 0.234 × 0.234 μm

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