Fig. 4: Time-resolved 3D qualitative and quantitative profiling of NP biokinetics in intact VAAD lungs enabled by LungVis 1.0.

a–p Longitudinal 3D view of NP distribution pattern in the left lung after ventilator-assisted aerosol delivery (VAAD) at time points of 0 h, 2 h, 24 h, and 14 d. a, e, i, m AI-powered visualization of NP airway deposition pattern. b, f, j, n Visualization of NP lung distribution (raw LSFM images). AI-powered global (c, g, k, o) and cellular resolution views (d, h, l, p, within white dashed lines) of NP acinar deposition pattern. Purple arrowheads refer to low NP deposition in parts of the upper lung after VAAD. Blue arrowheads indicate different NP (re-)distribution features in PAR. PB: Primary bronchus, TB: Terminal bronchioles. Scale bars: 1000 µm (a–c, e–g, i–k, m–o), 400 µm (ROIs), 200 µm (d, h, l, p). Representative data from n = 7 VAAD_0 h, n = 3 VAAD_2 h, n = 6 VAAD_24 h, n = 5 VAAD_14 days, independent biological replicates. NP intensity (brightness) optimized for spatial visualization, was scaled to match temporal lung homogenate dose trends but may not reflect the actual dose. q Schematic depiction of NP inter-acinar and intra-acinar transfer in the VAAD lungs (created partially with Biorender.com). r, s Longitudinal bronchial and acinar NP deposition fractions in the lung. n = 8 ITLI_0 h, n = 3 ITLI_2 h, n = 8 ITLI_24 h, n = 5 ITLI_14 days, n = 7 VAAD_0 h, n = 3 VAAD_2 h, n = 6 VAAD_24 h, n = 5 VAAD_14 days, independent biological replicates. t, u Fraction of NP retention in different compartments of the lung. The retention fractions are normalized to the mean dose at 0 h. n = 3, independent biological replicates. Data are presented as mean ± SD, calculated using one-way ANOVA with Holm-Šídák’s multiple comparison test (r–u). p values are indicated separately for ITLI (r) and VAAD (s) groups. Additional p values for bronchial dose fractions between ITLI_0 h and VAAD_0 h determined using unpaired two-tailed t-test (r). Source data are provided as a Source Data file.