Figure 3
From: Imaging multiple phases of neurodegeneration: a novel approach to assessing cell death in vivo
Identification of different patterns and phases of cell death in vivo. (a, b) Random sample (n=30) of single cells selected according to whether they were DiI positive (white spots, DiI retrogradely labelled retinal ganglion cells, a) and stained with annexin V-488, PI or both during time-lapse video (light blue spots, b). (c–p) Three patterns of fluorescent labelling were identified: ‘PI only’ (red spots, (c), and the representative cell, arrowed in (e–g) video stills with the corresponding signal intensity profile in (h)), ‘Annexin first’ (annexin-V positive and subsequently PI positive, green spots (c, i–l)), ‘similar profiles’ (positive labelling for both dyes at the same time, yellow spots, (c, m–p)). The mean times to peak intensity (d) revealed that the appearance of the ‘PI only’ cells was significantly quicker than either the ‘annexin first’ or the ‘similar profiles’ cells. Intensity profiles of annexin V and PI changed significantly over time (h, l, p). These results provide evidence that neuronal cells in vivo show considerable heterogeneity with regard to the kinetics of necrotic and apoptotic cell death, and also the relative timing of phosphatidylserine exposure (early phase) and nuclear condensation (late phase). Time points and scale bar 100 μm as indicated
