Fig. 1

a, Simplified scheme of the optical setup. b, In this setup, the light-sheet excitation and emission pass through a single objective. The fluorescence is collected by O₁ and relayed downstream with full NA detection, ensuring high-resolution imaging. c, The full NA detection is achieved by oblique remote focusing using a bespoke objective with a monolithic glass tip and zero working distance. The glass tip compresses the collection half-angle allowing a tilt range from 0 to 55°. d, During imaging, the stage moves the sample along the scanning axis. To avoid motion blur, the galvo mirror moves the light sheet alongside the stage movement during the camera exposure for each image. The galvo mirror moves back during the readout time and restarts this compensatory movement during the next exposure. Illumination and detection planes remain centered along the entire optical train to give optimal light collection, minimal aberrations and thus pristine image quality. e, Our instrument is capable of dual light-sheet excitation. This improves illumination coverage and image contrast, as for most points in the sample, one of the two light-sheet orientations will have a shorter penetration depth through the sample giving a more contrasted and complete image. The dual-view imaging is achieved through an imaging flipping module consisting of two galvo mirrors and three normal mirrors along the optical path (f and g). f, The illumination light goes along the path highlighted in orange or blue, resulting in opposing incident angle at the sample space. g, Similarly, the fluorescence light goes through either of the two paths, resulting in the flipping of the image with respect to that of the other path (blue and orange arrows before and after propagation through the unit), ensuring that the intermediate image is always formed on the front surface of O₃. h, The microscope is converted from upright (dipping, left side) to inverted (immersion, right side) by repositioning the coverslip from the focal space of O₂ to that of O₁, without sacrificing the optical performance.