Figure 5.

Workflow for mounting cleared tissue samples for lightsheet imaging

Samples can be mounted using agarose (A, C and D) or without agarose (B) for the Zeiss Lightsheet Z1 and Z7 systems with either a sample claw on the Z7 LSFM system (A-B) or a mounting screw on the Z1 and Z7 LSFM systems (C-D). For methods A and C, the tissue of interest and the mounting holder will be mounted in the solidifying agarose subsequently. For method B, the sample will be directly attached by a super glue. For method D, pre-heated agarose will be poured into a cryo-mold until 1 cm space remains at the top. The sample will be placed in the cryomold, chilled at 4°C, then removed from the cryomold and as much agarose as possible trimmed away to allow the sample to fit into the sample holder and decrease the amount of agarose the laser beam must pass through.