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. 2021 Feb 7;13(2):235. doi: 10.3390/pharmaceutics13020235

Figure 5.

Figure 5

NP uptake assays. (A) PLGA-PEG NP and PLGA-PEG-TFA NP uptake assay characterized using an Odyssey Imaging System. Both types of NP were incubated with C28/I2 cells for 2, 4, 8, 24, 48, or 72 h. (B) Confocal microscopy images of cell membrane stained with DiD (red) (644 nm excitation, emission filter 670/40), nucleus stained with DAPI (blue) (365 nm excitation, 40 ms, emission filter 450/50) and FITC NPs (green) (490 nm excitation, emission filter 515). Images taken after 24 h of cell incubation with NPs. The images were taken with a Leica SP5C Spectral Confocal Laser Scanning Microscope. (B1) Control sample (only cells with no NP treatment), (B2) cells and PLGA-PEG-TFA NPs, and (B3) cells and PLGA-PEG NPs. (C) Fluorescence microscopy images showing internalization of fluorescent PLGA-PEG and PLGA-PEG-TFA NPs. Visualization after 24 h of incubation with NPs in PBS. (C1) PLGA-PEG NPs, and (C2) PLGA-PEG-TFA NPs. Cell membrane stained with CD44-PE (red), cell nucleus with DAPI (blue) and NPs with NIR dye (green). The images were taken with a Leica fluorescence microscope at 43× magnification. Asterisks indicate significant difference in two-way ANOVA, ** p < 0.01 and **** p < 0.0001.