Figure 5.

Afterglow Luminescence Monitored ¹O₂ and Correlated with Immunogenic Cell Death and Anti-Cancer Efficiency In vitro. (A) Illustration of the afterglow monitoring photodynamic therapy. (B) Afterglow images of CT26 cancer cells incubated with different concentrations of SPN(NIR-3). (C) The corresponding afterglow intensity, quantified from (B). (D) The relative viability of CT26 cancer cells incubated with different concentrations of SPN(NIR-3), with 660 nm laser irradiation for 10 mins or not. (E) Correlation between afterglow intensity and cellular inhibition rate for SPN(NIR-3), calculated from (C) and (D). (F) Fluorescent confocal images of CT26 cancer cells treated with SPN(NIR-3) and 660 nm laser irradiation or not. The cell nuclei and ¹O₂ were detected using Hochest and SOSG, respectively. (G) Quantification of extracellular ATP concentration. (H) Correlation between afterglow intensity and the secreted ATP concentration, calculated using data from (C) and (G). (I-J) Fluorescent confocal images of CT26 cancer cells incubated with SPN(NIR-3) and 660 nm laser irradiation or not. The cell nuclei, CRT and HMGB1 were detected using Hochest, APC-conjugated anti-CRT antibody, and Alexa Fluor 594-conjugated anti-HMGB1 antibody respectively. (K) Flow cytometry analysis of CRT exposure for CT26 cancer cells treated with different concentrations of SPN(NIR-3) + irradiation. (L) Correlation between afterglow intensities and normalized CRT levels, calculated using data presented in (C) and (K).