Fig. 2: Qualitative and quantitative profiling of initial NP distribution in lung airways and acini enabled by LungVis 1.0.

a Schematic illustration delineates pulmonary NP delivery methodologies, longitudinal sampling, and quantitative NP dose in tissue homogenates and spatial NP dosimetry (created with Biorender.com). b Representative TEM and epi-fluorescence images of MF NPs. n = 3, independent replicates with similar results. Scale bars: 500 nm (left) and 15 µm (right). c Optical transparency of a mouse lung prior to and after tissue clearing. Scale bars: 1 cm. d–o AI-powered holistic mapping and cellular resolution views of NP lung distribution for four delivery routes stratified for the airway (e, h, k, n) and acinar region (f, i, l, o). d, g, j, m Schematic depiction of NP airway and acinar deposition (created with Biorender.com). Orange and white arrowheads indicate central/upper and lower/peripheral bronchioles, respectively. Blue arrowheads show NP deposition in upper peripheral regions (e, h). Red arrowheads show tracheal-deposited NPs (k). An acinus is delineated into the proximal and distal acinar region (PAR and DAR) artificially separated by black (d, g, j, m) or yellow (f, i, l, o) dashed lines. TB: terminal bronchioles. Scale bars: 1000 µm (overview), 100 µm (cellular resolution). p Quantitative analysis of NP-lung delivery efficiency. n = 5 INLA, n = 3 ITLI, n = 7 VAAD, n = 5 NOAI, biological replicates. q Central/peripheral NP deposition ratio. n = 3 INLA, n = 7 ITLI, n = 5 VAAD, n = 5 NOAI, biological replicates. r Bronchial/acinar NP deposition ratio. Representative data (d–o) from n = 3 INLA, n = 8 ITLI, n = 7 VAAD, n = 4 NOAI, biological replicates. s Linear correlation between C/P ratio and B/A deposition ratio with mean and 95% confidence intervals indicated by solid and dashed lines, respectively. n = 3 INLA, n = 5 ITLI, n = 6 VAAD, n = 4 NOAI, biological replicates. Data are presented as mean ± SD, calculated using one-way ANOVA with Holm-Šídák’s multiple comparison test (q, r). Source data are provided as a Source Data file.