Figure 4
From: Near infra-red labelling and tracking of corneal endothelial cells in-vivo
In vivo tracking of corneal endothelial cells. (a–c) The top row shows DIR stained rabbit corneal endothelial cells at day 3, 5 and 28 post wounding, imaged in vivo. (d–f) The middle row shows the same cells imaged with immunohistology at the same time points, after the animal was sacrificed. (g–i) The bottom row shows images acquired with conventional in vivo confocal microscopy. Each column represents images taken from the same cornea at the same time point (a) In vivo DIR imaging at day 3 showing migrating endothelial cells at the leading edge of the wound. Cells show an elongated morphology with incomplete monolayer formation. Marked variation in DIR uptake is seen between adjacent endothelial cells, with some staining intensely (b) At day 5, a complete monolayer is seen with high degrees of pleomorphism and broad intercellular junctions. (c) By day 28 cells have regained a more regular morphology and tight junctions have become narrower. Fixed cells stained with phalloidin, ZO-1 and Hoechst show similar morphological features to in vivo DIR imaging. (d) At day 3 cells are elongated with no junctional ZO-1 staining. (e) At day 5 a complete monolayer is seen with incomplete junctional ZO-1 staining and pleomorphic cells. (f) At day 28, regular cells with mature borders are seen. (g) Conventional in vivo confocal imaging failed to acquire useful images at day 3, and (h) day 5. (i) At day 28 a stable monolayer of cells was visible on conventional confocal microscopy, but nuclear morphology was more prominent than cell border morphology. Scale bar: 100 µm.
