Fig 2: Dynamic IR imaging of CSF tracer influx. (a) Schematic of experimental time course with representative images showing injection of dual tracers into the cisterna magna at (i) t=0 and dynamic in vivo imaging of IR tracer movement with (ii) a LICOR Pearl Trilogy imaging system through an intact skull, followed by surface imaging of both (iii) IR and fluorescent tracer, and (iv) subsequent imaging of fluorescent tracer and (v) immunofluorescence-probed slices. (b) Representative dynamic time series of IR LICOR images from mice anesthetized with 2% Iso or K/X acquired every 2 min from 20 to 40-min post-tracer infusion. (c) Representative image of the left and right dorsal cortical surface ROIs used for analysis of dynamic transcranial images and (d) quantification of the average cortical fluorescent intensity of dynamic images over time (20 to 40 min). Data are mean ± SEM from n=10 mice/group and were analyzed by repeated measures two-way ANOVA followed by Šidák’s post hoc test (p<0.01 for time points 1–5, p<0.001 for time points 6–12). (e) Representative dorsal and ventral LICOR whole-brain surface images taken following perfusion and removal of the brain from the skull. (f) Comparative analysis of average fluorescence intensity of IR tracer of LICOR dorsal and ventral ex vivo surface images of mice anesthetized with K/X or 2% Iso (**p<0.0019; ****p <0.0001), data are mean ± SEM from n=10 mice/group and were analyzed by two-way ANOVA followed by Šidák’s post hoc test.

Fig 3: Trial design and imaging workflow. (A) Panitumumab-IRDye800CW was infused in patients with contrast enhancing high-grade gliomas prior to surgery. (B) Intraoperative near-infrared imaging of the surgical field with neuronavigation (open field) and resected tissue (closed field). OR: operating room. (C) Microscopic and macroscopic NIR fluorescence imaging of fixed tissue and corresponding histopathological stainings post surgery. Red arrows: tissue imaging path; gray arrows: tissue processing path. NIR fluorescence imaging instruments (vendors): ? SPY-PHI (Novadaq); ? IGP-ELVIS (Li-COR Biosciences); ? Pearl Trilogy (Li-COR Biosciences); ? Odyssey CLx (Li-COR Biosciences). Images reproduced with permission from Medtronic, Stryker, and LI-COR, Inc.

Fig 4: Targeting zoledronic acid (ZA) using PAPC-L.a A diagram showing ZA encapsulated PAPC nanoliposomes (ZA-PAPC-L) depleting M2 TAMs. b Physicochemical properties of AS-HSPC-L and AS-PAPC-L and encapsulation efficiency (%EE). c % cell viability of THP-1 differentiated M1 and M2 macrophages 24 h after the treatment of free ZA, ZA-HSPC-L or ZA-PAPC-L (left to right: ***p = 0.000043, ***p = 0.000049). Data represents mean ± standard error of the mean (SEM) for three independent experiments. Statistical analysis was performed with Multiple unpaired t-tests with correction for multiple comparisons using the Holm–Sidak’s method. d Tumor growth curves in orthotopic 4T1 breast tumor model in mice. Both ZA-HSPC-L and ZA-PAPC-L were injected with dose equivalent to 0.5 mg/kg ZA, 100 µl injection volume, i.v., twice a week, when tumors became ±100 mm3. Before sacrifice, mice were injected with 2.5 mg of D-luciferin and were imaged after 10 min to detect bioluminescence signal using Pearl Trilogy imager (LICOR) (left to right: *p = 0.049, *p = 0.034, *p = 0.014, *p = 0.018 between Vehicle and ZA-PAPC-L). Mean + SEM, N = 6 mice per group, Statistical analysis was performed with unpaired two-tailed Student’s t-test. e Representative images showing the isolated tumors with bioluminescence signal (green). f Graphs showing the tumor weight of the isolated tumors at the end of the experiment. N = 6 mice per group (*p = 0.043). Statistical analysis was performed with unpaired two-tailed Student’s t-test. g Box graphs showing quantitative real-time PCR results showing the effect of ZA-HSPC-L and ZA-PAPC-L on different markers for pre-metastatic niche in lungs. N = 6 mice per group (left to right, Ym1: *p = 0.010, **p = 0.0043, *p = 0.044; Ccr2: *p = 0.018, *p = 0.042; Postn: **p = 0.0020, **p = 0.0031, *p = 0.021; Col1a1: **p = 0.0088, **p = 0.0011, *p = 0.048; Sdf-1: *p = 0.014, **p = 0.0079, *p = 0.012; iNos: ns = non-significant). Data in (f and g) represent the box-and-whisker plot in which every dot represents a data point and the line in the box as median; the box shows the upper to the lower quartiles; the whiskers show the minimum to the maximum value. Statistical analysis was performed with unpaired two-tailed Student’s t-test. Source data are provided as a Source Data file.

Fig 5: Real-time surgical specimen analysis. Fluorescence-guided analysis of the mucosal tumor and of the deep resection margins in HNSCC patients was performed using Li-COR PEARL Trilogy. a Representative image of a mucosal tongue tumor with an insufficient superficial surgical margin (2 mm). b Representative image of a mucosal tongue tumor with a sufficient superficial surgical margin (> 5 mm). c Representative image of a deep surgical resection margin negative for fluorescence which correlated with a tumor-negative margin. d Representative image of a positive fluorescent lesion on a deep resection margin correlating with a tumor-positive surgical margin.