Figure 3:

Schematic representation of the culturing and treatment of heterocellular Panc (PDAC-PCAF) spheroids followed by an imaging-based analysis of treatment response.

A comprehensive image analysis procedure for structurally complex organotypic cultures was used for the quantitation of fractional viability of spheroids following NIR photodynamic activation (A) using TR-PINs and PSNs (B). The NIR photodynamic activation regimen used was 690 nm light irradiation with 25 or 50 or 75 or 100 J/cm² at 150 mW/cm². Quantitation of fractional viability of MIA PaCa-2 and PCAFs spheroids following NIR photodynamic activation (25, 50, 75, 100 J/cm²) at a log₁₀ [BPD-PC] (nM) = 2.3 (250 nM of BPD-PC equivalent) using TR-PINs (C). Viability heatmap images of heterocellular (PDAC + PCAF) spheroids following NIR photodynamic activation of TR-PINs with increasing concentrations of the photosensitizer BPD-PC (D). Quantitation of fractional viability of spheroids following NIR photodynamic activation (100 J/cm²) using TR-PINs (E). (mean ± SEM; n = 9–12 for b–e; one-way ANOVA with a Tukey post-test; ****P ≤ 0.0001, ***P ≤ 0.001, **P ≤ 0.01).