At the top is the optical layout of the detection path of the eSPIM simulated with Zemax. O1–O3: objective lenses—respectively, water immersion 60×/1.27-NA, air 100×/0.9-NA and water immersion 60×/1.0-NA. The objectives are modeled using parameters that were inferred from patent literature of similar objectives from the same producer: 60×/1.27-NA water-immersion objective from patent US8199408, 100×/0.9-NA air objective from patent US9341832, and 60×/1.0-NA water-immersion objective from patent US7133212. L1–L5: achromatic doublet lenses, with focal lenses as 200 mm, 75 mm, 125 mm, 150 mm and 125 mm. The parameters of those lenses are obtained from the Thorlabs website. At the bottom, the Huygens PSFs (image in the camera/detector plane) are given for five different position (i–v) in the focal space of O1. The FWHMs along the x- and y-axes are, respectively, 255 ± 2 nm and 355 ± 5 nm (average of the measurement from the five PSFs, mean ± s.d.).