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normoxia with the 808 nm laser, as proven by the widespread red fluorescence color, and penetrated deep into the spheroids (10–80 μm Z-track images from slices with a 10-μm step size). However, a negligible cytotoxicity was shown in the spheroids treated with UCNPs@SiO2-NH2@FA/PEG/TCPP in hypoxia with the 808 nm laser and in normoxia without the 808 nm laser (Figure 8B). This finding Pinhadrimcacteeudticsth20a2t0U, 1C2, N11P02s@SiO2-NH2@FA/PEG/TCPP would presumably have the practical poten1t2iaolf t1o7 have a photodynamic antitumor efficacy for the relevant tumor in vivo. Figure 7. Live/Dead assay of PBS, free TCPP, UCNP@SiO2@FA/PEG, and UCNP@SiO2@FA/PEG/TCPP Figure 7. Live/Dead assay of PBS, free TCPP, UCNP@SiO2@FA/PEG, and UCNP@SiO2@FA/PEG/TCPP under normoxic or hypoxic conditions with or without laser irradiation (660 nm, 30 min, 10 mW/cm2 under normoxic or hypoxic conditions with or without laser irradiation (660 nm, 30 min, 10 mW/cm 2 for free TCPP; 808 nm, 30 min, 1.5 W/cm2 for others). Pharmfaocreuftriecse2T0C20P,P12;,8x08FOnRmP,E3E0RmRiEnV,1IE.5WW/cm2forothers). 12of16 Figure 8. (A) Images for the Live/Dead assay in 3D multicellular HeLa cell spheroids under the Figure 8. (A) Images for the Live/Dead assay in 3D multicellular HeLa cell spheroids under the 22 predetermined conditions (808 nm, 30 min, 1.5 W/cm ). (B) Z-stack images of six 10–20-μm slices in predetermined conditions (808 nm, 30 min, 1.5 W/cm ). (B) Z-stack images of six 10–20-μm slices in the the HeLa cell spheroids. HeLa cell spheroids. 3.7. Cellular Uptake of UCNPs@SiO2-NH2@FA/PEG/TCPP into HeLa Cells The cellular uptake of UCNP conjugates into HeLa cancer cells was visualized by Bio-TEM and CLSM. As shown in Figure 9A, UCNPs@SiO2-NH2@FA/PEG/TCPP was internalized into the cells in 10 min (red fluorescence for conjugated TCPP). This result indicates that the UCNPs are taken up by the cells, presumably due to folic acid-mediated endocytosis and small size-based nonspecific uptake [55]. To confirm the internalization of UCNPs@SiO2-NH2@FA/PEG/TCPP in HeLa cells, cells were photographed by Bio-TEM (Figure 9B). The diameter of HeLa cells was estimated to be 20–40 μm,PDF Image | Red LED Erbium Upconverting Nanoparticles v Cervical Cancer
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