M-Cube — towards correlative multi-scale light-sheet imaging using a compact, modular and moving illuminator

Abstract

The crossed arrangement of excitation and collection optics is the defining feature of selective plane illumination microscopes. It results in an axial sectioning given by the thickness of the light sheet, whereas the lateral resolution depends on the numerical aperture of the collection optics. One disadvantage of this optical scheme is that it has been difficult to image large fields-of-view at high spatial resolution. Yet, it is often not necessary to image the entire sample at high resolution. Instead, a zoom from mm-scale over-views to regions-of-interest that are imaged at μm is often sufficient, e.g., for studying neuronal networks that simultaneously comprise cm-long connections and tiny (sub-μm) synaptic contacts, spanning 6 orders of magnitude. Observations over such different spatial scales typically require the use of different instruments. We here describe our ongoing efforts to build and characterise a compact light-sheet module designed for correlative micro- meso- and macroscopic imaging. Its particularity is that moves with the sample between micro- and macroscopic imaging arms. Based on a compact illuminator and flexible sample carrier the module is mounted on a motorised long-range, high-precision microscope table. The ability to perform seamless back-and-forth multi-scale imaging comes from a strict registering of image coordinates.