Fig. 1 The method used to tile LLSs in typical LLS microscopes. (a) A simplified optical configuration of the LLS microscope illumination path. (b) The amplitude and phase profile at the center cross section of a simulated optical lattice, and the corresponding binary phase map acquired from either of them to generate the optical lattice in LLS microscopes. (c) The amplitude and phase profile at an off-center cross section of the same simulated optical lattice in (b), and the corresponding binary phase map acquired from either of them to tile the optical lattice in LLS microscopes. (d-f) 3D volume renderings of simulated optical lattices generated by modulating the illumination light of the given amplitude profile using the given phase profiles at the image plane where the SLM locates in the LLS microscope. The white frames indicate the detection FOV.

Fig. 2 Simulation results of tiling optical lattices generated by applying binary phase maps to the binary SLM used in typical LLS microscopes. (a-d) Four binary phase maps acquired from different off-center cross sections of the same optical lattice (NA_OD = 0.4, NA_ID = 0.22) used to tile the optical lattice, and the intensity profiles of the corresponding tiling optical lattices and LLSs at the indicated cross sections. NA_OD and NA_ID are the illumination numerical apertures (NA) corresponding to the outer and inner diameter of the transparent optical annulus used to create the optical lattice. Scales bars, 5 µm in (a) XZ panel, and 10 µm in (a) YZ panel.

Fig. 3 The tiling distance limitation caused by the illumination light beam width. (a) A binary phase map applied to the binary SLM to modulate the illumination light with different beam widths of amp1 to amp3. (b-d) The intensity profiles of the tiling optical lattices and the indicated cross sections obtained by modulating the illuminating light of different beam widths with the same binary phase map shown in (a), demonstrating the tiling distance limitation caused by beam width of the illumination light. Scales bars, 5 µm in (b) XZ panel, and 10 µm in (b) YZ panel.

Fig. 4 Schematic diagram of the experimental lattice light sheet microscope.

Fig. 5 Characterization of tiling LLSs on a LLS microscope. (a) Images of an optical lattice tiled in dye solution. Tile 1 represents the regular LLS. (b) XY and YZ maximum intensity projections of a group of fluorescent particles embedded in agarose gel imaged using the tiling LLSs in (a), and zoom-in views of the marked areas. Scale bars, 10 µm. 1 µm in zoom-in views.

Fig. 6 An expanded mitotic cell imaged using tiling LLSs on a LLS microscope. (a) XY and YZ maximum intensity projections of the cell imaged with a regular LLS, and zoom-in views of the marked areas. (b-c) XY and YZ maximum intensity projections of the expanded cell imaged using tiling LLSs at different tiling positions, and zoom-in views of the marked areas. The arrows indicate the waist positions of the tiling LLSs. (d) XY and YZ maximum intensity projections of the reconstruction result, and zoom-in views of the selected areas. (e) XY and YZ maximum intensity projections of the deconvolved reconstruction result, and zoom-in views of the marked areas. Scale bars, 20 µm.