File:Stage-Specific-Changes-in-the-Water-Na+-Cl--and-K+-Contents-of-Organelles-during-Apoptosis-pone.0148727.s008.ogv

English: Long-term imaging of nuclear modifications induced during 500 ng/mL AMD treatment and subsequent recovery after AMD washing out studied by time-lapse confocal imaging of HeLa cells stably expressing H2B-GFP. Time-lapse confocal imaging of H2B-GFP was performed by two-photon excitation, every 5 minutes. Z-stacks of 85 optical sections were processed to obtain 3D images of chromatin and were assembled to produce one movie demonstrating the successive steps of 3D chromatin reorganization. Cells in exactly the same area were imaged during successive periods and conditions. During the first period of time (0 to 7h), cells were maintained in contact with AMD to induce apoptosis. At the end of this period, 7 cells (on a total of 34 cells) were engaged in apoptosis. Cells were then rinsed several times in PBS and fresh culture medium to wash out AMD. During the second period (7h45 to 22h 16 minutes), cells were maintained in culture medium without AMD and the same area was imaged. At the end of this period, 22 cells were engaged in apoptosis and 12 were not engaged in apoptosis (unaffected cells or stage 0 cells). Culture medium without AMD was changed each day during 3 days and then cells in the same field of view were imaged during 6 hours and 25 minutes. During this period, 10 cells were imaged: none engaged in apoptosis and five entered mitosis.