Design and Build of a Customizable Light Sheet for Visualization of Cytoskeleton Networks

Abstract

Reconstituted cytoskeleton composites have emerged as a valuable model system for studying non equilibrium soft matter. Faithful capture of the dynamics of these three dimensional, dense networks calls for optical sectioning often associated with fluorescence confocal microscopes. However, recent developments in light sheet microscopy has established it as a cost effective, and at times superior, alternative. To make the LSM accessible to cytoskeleton researchers less familiar with optics, we present a step by step beginners guide to building a versatile light sheet microscope from off the shelf components. To enable sample mounting with traditional slide samples, our LSM follows the single objective light sheet design, which utilizes a single objective for both excitation and emission collection. We describe the function of each component of the SOLS in sufficient detail to allow readers to modify the instrumentation and design to fit their specific needs. Finally, we demonstrate the use of our custom SOLS instrument by visualizing asters in kinesin driven microtubule networks.