Figure 1.

Nanoneedle interfacing with human corneal endothelial cells in vitro. A) Schematic representation of the nanoinjection approach for cultured primary HCEnCs. Image created with Biorender.com. B) Confocal microscopy orthogonal projections of nanoneedles (FITC labeled, green) interfaced with the cytosol, outlined by ZO‐1, and the nucleus of HCEnCs. Nanoneedles colocalize with HCEnCs. ZO‐1 staining (red) with DAPI (blue) nuclear counterstain. Scale bar 20 µm. Images were obtained immediately after nanoneedle assisted interfacing by centrifugation. C) Phase‐contrast image of the primary HCEnCs culture showing retained morphology following nanoneedle interfacing (nN), similar to the untreated control (ctr). Images were obtained immediately after the deinterfacing. D) Immunofluorescence microscopy of HCEnCs showing retained hexagonal morphology and ZO‐1 marker upon nN interfacing (nN) as well as in untreated HCEnCs (ctr). ZO‐1 staining (red) with DAPI (blue) nuclear counterstain. Scale bar 50 µm. Images were obtained 72 h following nanoneedles interfacing. E) Immunofluorescence microscopy of Caspase 3/7 activation. Lack of nuclear staining with faint cytoplasmic staining 72 h following nanoneedle interfacing (nN), comparable to untreated control (ctr) demonstrate lack of Caspase 3/7 activation, indicating absence of apoptotic events. Caspase 3/7 (green) staining with DAPI (blue) nuclear counterstain. Scale bar 50 µm.