In the tumor of the treated animal, an increasing deviation between the measurements and the fitted curves was observed from day 2 onwards, between 500 and 800 nm. This indicates that fluorophores other than the ones included in the standard fit model (collagen, elastin, NADH, and FAD) were

measured. This additional fluorescence activity buy Panobinostat (from now on called fluorescence residual) was seen in all the treated tumors at days 4 and 7. The longitudinal kinetics for each model-fitted AFS parameter and the calculated fluorescence residual across all treated and control animals are shown in Figure 4. The plotted linear trend for the fluorescence residual in tumor was significantly different between the treated and the control groups (P = .018). No significant trends were observed for the total fluorescence intensity, collagen + elastin, and the optical redox ratio. Figure 5 shows the longitudinal check details changes of the fluorescence residual in tumor, liver,

and muscle across all animals from both groups. The additional fluorescence is not present in muscle and liver tissues, indicating a tumor-specific effect. In an attempt to better understand the origin of the additional autofluorescent emission (mainly above 600 nm) seen in the treated animals, two-photon confocal fluorescence microscopy images recorded in a spectral range of 600 to 700 nm were compared with adjacent tissue sections that were stained with HE (Figure 6). The samples were collected after 1 week of follow-up, i.e., when the differences seen in AFS signals were maximal. In the treated tumor samples, numerous fluorescent foci were present. These foci correlated with cellular structures rather than with collagen deposits or necrotic areas. It remains to be determined

whether this specific fluorescence originated from stromal or tumor cells. Ixazomib clinical trial For the two-photon images recorded in the spectral ranges 400 to 500 nm and 500 to 600 nm, no considerable differences were seen when comparing both groups. The evaluation of pathologic response of tumors to cisplatin using various histologic dyes and immunohistochemical biomarkers is illustrated in Figure 7. A strong increase in nuclear DNA damage was seen 24 hours after cisplatin administration using γ-H2AX as a marker. From day 2 onwards, a significant decrease in the proliferation marker Ki-67 and an increase in apoptosis-related cell death (CC3 marker) were observed. Analysis of MT-stained slides showed increased amounts of fibrotic tissue 4 to 7 days after treatment that corresponded to the HE images. An increase in lipids (Oil Red O) was seen over time. In Figure 8, A and B, fractions of vital, necrotic, and fibrotic tumor tissues for both groups are shown as quantified on the HE-stained tissue slides.