From OpenSPIM to FlowSPIM: Enhancing the versatility of a light-sheet microscope through an iterative design process

Abstract

Light-sheet microscopy is ideal for the 3D imaging of large biological specimens, yet systematic documentation on adapting microscope designs to diverse experimental requirements remains limited. We present the iterative process used to design and build a modular light-sheet microscope and adapt it to different sample types and experimental conditions, including live and fixed zebrafish, Drosophila, and mouse gastruloids, as well as cellulose nanocrystals. Starting from a stripped-down OpenSPIM platform, we gradually made our microscope more versatile by adding hardware such as additional laser channels and optical filters, and new protocols for sample handling. In its final iteration, FlowSPIM is a new flow-based incubation technique for 3D tissue culture.

arXiv commentElectronic material is available at https://doi.org/10.5281/zenodo.23898349 . The manuscript has 13 figures and 2 tables. This is a revised version after peer-review. Updates include product numbers and vendors of all the parts, ray diagrams, details of the optical configurations, and a new Discussion chapter with explicit troubleshooting instructions