Designing a Singular Objective Light-Sheet (SOLS) Microscope for Visualization and Characterization of Cytoskeleton Networks*

Light-sheet fluorescence microscopy (LSFM) has become a popular method for high resolution imaging of thicker (~>100??m) samples in biological research. Held in comparison to other high resolution imaging techniques, such as confocal microscopy, LSFM is notable for its low amount of photobleaching which increases the timescales over which samples can be imaged. In particular, one type of LSFM build termed a Single-Objective Light-Sheet (SOLS) has provided the ability to image samples mounted using traditional top down microscope slides, which was not possible in previous LSFM builds. The applications of this type of microscope are great in number, yet the build and alignment process of a SOLS microscope remains a complicated process, even for a user with a background in optical systems. Here, we present a simplified and reliable procedure for the construction of a modular SOLS lab microscope, thereby making this process accessible to researchers with minimal prior optics experience. We provide discussions on the advantages and disadvantages of using particular optical components over others, then design a step-by-step guide on how to construct and align a SOLS microscope using commercially available parts. After the build, we demonstrate the capabilities of the system by imaging in 3D reconstituted cytoskeleton networks and discuss future capabilities of similar systems.