Figure 7: Pharmacokinetics of DY-635[NP](−).

(a) Time course of arterial plasma concentration after central venous application of DY-635[NP](−) (left ordinate) at time ‘0’ and appearance of the desorbed dye in bile (right ordinate) (n=3, mean±s.e.m). (b) Percentage of recovered free dye in bile and urine after administration of DY-635[NP](−). (c) Fluorescence decay over liver parenchyma in the Cy5 channel after administration of unbound DY-635 or DY-635[NP](−) revealing exponential decay for the free dye and almost linear decay for DY-635[NP](−)-associated fluorescence (n=3 per 25 ROIs per n; mean±s.e.m.). (d) Intravital epifluorescence microscopy to visualise acinar distribution of DY-635[NP](−). Upper panel background fluorescence of the liver (blue). Middle panel: Heatmap (blue: low intensity, red: high intensity) reflecting signal distribution which is associated with the pericentral region of the liver lobule early upon injection of DY-635[NP](−) and spreads towards midzonal and even periportal region over time. Lower panel: corresponding false colour images of association of DY-635-associated fluorescence (red) with liver parenchyma (green): fluorescence is restricted to the vascular compartment early upon injection (5 min); yellow signal indicating colocalisation of dye with liver parenchyma is increasingly observed over time. (scale bars 100 μm). (e) Theranostic use of the free dye DY-635 to predict subsequent uptake of DY-635[NP](−). DY-635 was injected at time ‘0’ followed by injection of DY-635[NP](−) at 20 min. (f) Intensity of fluorescence signal over liver parenchyma; right panel: signals obtained upon injection of free dye and dye-functionalised nanoparticles are correlated. (n=2, 20 ROIs per n; mean±s.e.m.) (scale bar 100 μm).