Figure 4

In vivo examination of mitochondrial motility and membrane potential in stage M1-M4 mitochondria within a single areole of a window stage lace plant leaf. Still images selected from time-lapse videos at time 0 and time 30 seconds following CMXRos staining. Mitochondria in time 30 sec images have been false colored green to allow for comparative overlay images to demonstrate mitochondrial motility. A, D, G and J) time 0 seconds CMXRos stained images of M1, M2, M3 and M4 mitochondria over the PCD gradient (NPCD-LPCD), respectively. B, E, H and K) time 30 seconds CMXRos stained images of M1, M2, M3 and M4 mitochondria over the PCD gradient (NPCD-LPCD), respectively. C, F, I and L) Overlay of time 0 and 30 second still images of M1, M2, M3 and M4 mitochondria over the PCD gradient (NPCD-LPCD), respectively. Note that when mitochondria have not moved, overlay images appear yellow. These overlay images characterize the rapid mitochondrial movement of M1 and M2 stage mitochondria, followed by the decrease in mitochondrial motility in M3 and M4 stage mitochondria. Also note the loss of mitochondrial staining in M4 mitochondria, indicating these organelles appear to have undergone a membrane permeability transition and have lost their membrane potential. Still images A, B and C taken from additional file 5. Still images D, E and F taken from additional file 6. Still images G, H and I taken from additional file 7. Still images J, K and L taken from additional file 8. Scale bars (A-I) = 10 μm.