The second Systematic Live Imaging Collection of Embryogenesis (SLICE-2) is a collection of sixteen isotropic 3D fluorescence live imaging datasets documenting gastrulation and early germband elongation in the red flour beetle Tribolium castaneum. Imaging was performed using light sheet fluorescence microscopy (LSFM) in combination with the transgenic AGOC{ATub’H2A/H2B°NB-mEmerald} #1 subline, which expresses mEmerald-labeled nanobodies against histone H2A/H2B under the control of the constitutively and ubiquitously active tubulin alpha 1-like protein promoter. Each embryo was recorded in a single fluorescence channel along four directions spaced 90° apart. Recordings comprised up to 51 time points at 30 min intervals, corresponding to a total imaging duration of up to 25 h. The resulting image stacks were subjected to multiview fusion to generate isotropic 3D reconstructions of the data, which are provided as two different derivatives: weighted-average fusions and fusion-deconvolutions. Each dataset comprises one recording of one embryo and was processed using multiple image-processing branches. For convenience, the data are distributed across multiple archive parts corresponding to these branches: the basic branch (B) – part 1 the weighted-average fusion (F) and fusion-deconvolution (D) branches – part 2 the preliminary processing branch (P) – part 3 (and optionally parts 4–6 for selected datasets) Entries belonging to the same dataset are clearly labeled in the entry title and cross-linked in the ‘Additional details’ section. Each entry also includes comprehensive experimental metadata in the form of a human- and machine-readable XLSX file. In addition, the first entry of each dataset contains intensity-adjusted z projection t stack montage TIFF files together with corresponding AVI files for rapid inspection of the dataset in Fiji or any standard movie player. Detailed descriptions of the experimental procedures, image-processing workflow, folder structure, and file organization are provided in the associated Data Descriptor.