Beyond single cells: Ten years of commercial lattice light-sheet microscopy

Lattice light-sheet microscopy (LLSM) enables gentle, rapid, volumetric fluorescence imaging of living biological systems. The technique employs Bessel beam interference to produce a light-sheet which retains its thickness over a relatively large field-of-view compared to other light-sheets. In the decade since its commercialisation, LLSM has become an important tool for imaging dynamic cellular processes, while increasingly being applied to multicellular specimens including organoids, embryos, tissue explants and small model organisms. This primer introduces the optical principles and acquisition geometry of LLSM, with particular emphasis on commercial systems and the computational processing required to transform tilted stage-scan data into biologically interpretable image volumes. We discuss the strengths of LLSM for long-term, multicolour imaging, as well as its integration with advanced modalities including structured illumination and single-molecule localisation approaches. Focusing on applications beyond single cells, we examine practical considerations that determine experimental success, including sample suitability and preparation, acquisition strategy, and data handling. We further highlight the substantial computational challenges associated with LLSM, and open source tools available to support processing and analysis.