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. 2015 Jul 8;5:12175. doi: 10.1038/srep12175

Figure 4. Differences in efficacy of PDT between 2D monolayer cell cultures and 3D spheres.

Figure 4

(a-d) Images of T47D 2D cell cultures after PDT with LIVE (green) /DEAD (red) staining. 10 μM methylene blue in PBS was used as photosensitizer with light exposure doses of (a) 0 minutes, (b) 10 minutes, (c) 30 minutes, and (d) 60 minutes. The fluence per minute is 0.73 J/cm2. (scale bar: 50 μm) (e-h) Images of large (average diameter: 155 μm) T47D spheres after PDT with LIVE (green) /DEAD (red) staining. Light exposure doses of (e) 0 minutes, (f) 10 minutes, (g) 30 minutes, and (h) 60 minutes were used. (scale bar: 50 μm) (i-l) Images of small (average diameter: 95 μm) T47D spheres after PDT with LIVE (green) /DEAD (red) staining. Light exposure doses of (i) 0 minutes, (j) 10 minutes, (k) 30 minutes, and (l) 60 minutes were used. (scale bar: 50 μm) (m) Differences in PDT efficacy between T47D spheres and 2D monolayer culture with different light doses (fluence). The IF50 (half maximal inhibitory fluence) of 2D cell monolayer culture is estimated to be 4.9 J/cm2 (N = 5 cultures), while the IF50 of large spheres is estimated to be 17.8 J/cm2 (N = 10 spheres) (n) PDT efficacy differences as a function of fluence for large and small spheres. The IF50 of small spheres (average diameter: 95 μm) is estimated to be 9.4 J/cm2. (N =  10 spheres) The IF50 of large spheres (average diameter: 155 μm) is estimated to be 17.8 J/cm2. (N = 10 spheres) * refers to P < 0.05, and ** refers to P < 0.01.