Cells (2 × 105/ml) were incubated with GO at 10 mg/ml for 3 hours. For confocal imaging, cells were fixed in 3.7% paraformaldehyde for 30 min, and the cell membrane was stained with a lipid probe, DiO (green). The GO-membrane interaction was initially imaged using confocal laser scanning microscopy (Leica TCS SP8) and further via HyVolution. Taking advantage of the intrinsic photoluminescence of GO in the near-infrared (NIR) region, the sample were excited at a 633-nm laser and monitored at an emission spectrum larger than a 700-nm wavelength during confocal imaging. The GO signal was marked by red.

For TEM imaging, cells were collected and fixed in 2.5% glutaraldehyde [pH 7.4, in phosphate-buffered saline (PBS)] for 1 hour at room temperature to maintain the ultrastructure of the cells as close to the living material. Following that, the samples were dehydrated through an ethanol series, passed through a propylene oxide, and then embedded in a liquid resin. Subsequently, the resin block was sectioned by a microtome (Lecia EM UC6), and the sections were collected and stained with electron-dense strains (1% lead citrate following 4% uranyl acetate) before observation. Further evidence of the GO-membrane sandwiched structure was acquired under a Hitachi H-7650B microscope.

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