FRAP experiment on a C. elegans nucleus expressing GFP::Ce-lamin containing an R64P mutation

The movie was filmed with an FV-1000 confocal microscope (Olympus) equipped with an IX81 inverted microscope and a 60X/1.4 oil-immersion objective. Worms expressing the R64P GFP::Ce-lamin protein were photobleached in defined regions of specific nuclei by a 405-nm laser and were imaged with a 488-nm laser line for excitation and a 505-525-nm filter for emission. Fifteen frames were taken before the photobleach, two frames were taken during the photobleach, and 200 frames were taken after the photobleach. Frames were taken every 1.107 sec (Wiesel N, Mattout A, Melcer S, Melamed-Book N, Herrmann H, Medalia O, Aebi U, Gruenbaum Y. 2008. Laminopathic mutations interfere with the assembly, localization, and dynamics of nuclear lamins. Proc Natl Acad Sci 105: 180-185). Supplemental movie from Live Cell Imaging, 2nd ed., Cold Spring Harbor Laboratory Press.