Comparison of tumor background ratio in inflated and deflated lung.

<p>Boxplot demonstrating a statistically significant increase in tumor-background ratio with inflated lung compared to deflated lung, n = 7 (p = 0.003).</p

Representative CT axial images of mouse lung.

<p>(A) CT axial images of mouse lung with no CF800 injection (left) and at 48h post-CF800 injection (right). The arrows point to the location of the lung tumor. Both images are displayed at the same window and level. Note the higher CT signal observed in the tumor of the mouse injected with the CF800 agent. (B) Bulk tumor analysis demonstrating CF800 injected mice have a higher tumor signal than non-injected mice, p = 0.0002.</p

Correlation between NIR intensity and NIR mean fluorescence.

<p>Positive correlation between NIR intensity and NIR mean fluorescence. SPY can be used as a quantitative tool to accurately measure ICG liposome accumulation. Positive correlation (R² = 0.81) exists between measured SPY fluorescence image signal and average NIR intensity of tumor on histological section.</p

Experiment model.

<p>(A) Method of establishing orthotopic human non-small cell lung (H460) cancer house model. (B) Two representative examples of NIR ex-vivo imaging after injection with CF800.</p

Histology based characterization.

<p>Histology of tumor and normal lung compares boxplots of (A) vessel density p = 0.0037 (A), and (B) macrophage density p = 0.0041 (B) student’s t-test demonstrating they are lower in tumor compared to lung. However (C) NIR positivity is overall slightly increased and more variable in tumor. Correlation between vessel density and NIR positivity (D) is strongly negative whereas macrophage density (E) is not correlated to NIR positivity.</p

CF800 localization in tumor.

<p>(A) Histological section of tumor viewed with Tissuescope under NIR, (B) (H&E), (C) overlay of NIR, and immunocytochemical markers CD31 and F4/80. Areas of NIR signal detected with Tissuescope correlates with malignant cells on H&E staining. Malignant cells outlined in yellow.</p

Injection volume was optimized using <i>ex vivo</i> porcine lungs.

<p>(A) The lung cross sections showed the dimensions of injected ICG (100 μg/ml, paired with 5%BSA) which depended on the volume of the injection. Scale bars show 10 mm. (B) ICG injection of 1,000 μL resulted in significantly larger ICG florescence area compared to the other 2 doses (*p = 0.0033, Kruskal-Wallis). Each box consists of both sides of cross sections of 3 lungs (n = 6).</p

Multiple factors determine the ICG fluorescence intensity.

<p>(A) Autologous porcine plasma (PL) and 5% bovine serum albumin (BSA) enhanced the ICG fluorescence intensity, with the brightest intensity at 10 μg/ml (at 3cm). Top: normal images, middle: merged images, bottom: fluorescence images. (B, C) Fluorescence intensity depended on the distance from the objects. ICG was paired with PL.</p

Successful sentinel lymph node detection using the near-infrared fluorescence imaging.

<p>One of the subcarinal lymph nodes was successfully identified as a sentinel lymph node immediately after transpleural ICG injection (100 μg/ml, 100 μL) into the right lower lobe. The afferent lymph vessel was clearly visualized as well, showing a high contrast when compared to the background. Left: merged images, Right: fluorescence images.</p

Successful sentinel lymph node identification after pre-operative tansbronchial ICG injection.

<p>Electro-magnetic navigational bronchoscopy-guided transbronchial ICG injection (100 μg/ml, 100 μL) into the left lower lobe enabled the SN node identification by the NIR thoracoscope. (A) The left paratracheal lymph node was recognized as the SN. Left top: intraoperative NIR image, left bottom: <i>ex vivo</i> examination of the SN, right: <i>ex vivo</i> examination of the lymph node cross sections which showed select bright portions. Asterisk shows hemi-azygos vein. (B) The navigation guidance led the bronchoscope to the target accurately. The scale bar shows 10 mm.</p