DIY MuVi-SPIM Light-Sheet

This microscope is a multi-view selective illumination (MuVi-SPIM) microscope.  Two parallel light sheets are generated by Nikon 10x .3NA objectives.  Thickness of each light sheet is 1um.  Two opposing detection arms use Nikon 25X 1.1NA objectives and Hamamatsu Orca Flash 4.0 V2 Digital CMOS cameras.   The sample is bottom loaded into a water filled chamber, and movement of the sample is driven by an XY and Theta piezo motors.  The maximum imaging volume while using the 25X objectives is 512um *512um *1500um.  Within a single time point, the sample volume is imaged from one orientation using both cameras, and then the sample is rotated 90 degrees and the same volume is captured again.  These two perpendicularly oriented volumes are then fused using an in-house data reconstruction algorithms.  The output of this reconstruction is a single fused volume that includes information captured with both cameras from both orientations.

Best Application/s:  This microscope has been optimized for Drosophila larvae.  We are also planning on coupling  a DMD to this system to add optogenetic capabilities.