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Fig. 2 | BMC Biology

Fig. 2

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

Fig. 2

LSTM microscopy implementation. a LSTM optical path. Two symmetric light sheets are generated by using a cylindrical lens (CL), scan lens (SL), tube lens (TL), and illumination objectives. The galvo scanners are used to translate the light sheets perpendicular to their propagation direction, and the electrically tunable lens (ETL) for translating the thinnest part of the light sheets along the propagation direction. An input beam of ~ 10 mm diameter is then trimmed through an iris. A slit is placed after the ETL to control the effective numerical aperture of the illumination. An additional iris is placed between the SL and TL to control the light sheet width. The illumination axes are arranged at ~ 60° to the detection axis. A custom 3D-printed cap with a quartz coverslip is attached to the illumination objective to allow dipping in the immersion oil to ensure that the low NA illumination rays from an air illumination objective enter perpendicularly to the oil. The detection arm consists of a detection objective (Olympus 10×/0.6NA/8mmWD or 25×/1.0NA/8mmWD, both with correction collars), a tube lens, and an sCMOS camera. b 3D model of the LSTM microscope. A vertical breadboard was used to mount the caged optical assemblies via x-y manual translation stages to allow fine adjustments. A sample chamber was attached to a 3-axis (x, y, z) motorized stage assembly. See also Additional files 1, 2, 3, and Additional file 4: Video 1 for further details and Table 1 for complete parts list

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