bioimagesUnknownHeidi N. FridolfssonThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201004118JCB

DIC images from a representative time-lapse sequence of klc-2(km11) nuclear migration in hyp7 precursors. Cellular boundaries appear as depressions in the hypodermis and nuclei are exclusions of cytoplasmic granules. Embryos were dissected from gravid hermaphrodites and mounted on a 2% agar pad in M9 solution. Time-lapse microscopy of nuclear migration was observed using DIC optics and a 63x PLAN APO 1.40 NA objective on a Leica DM 6000 compound microscope. Images were acquired using a Leica DC350 FX camera and the LAS AF software (Leica) at a rate of 15 sec/frame.

Microtubules polarize in the direction of nuclear migration as imaged by EBP-1::GFP. Time-lapse microscopy of EB1::GFP expressing embryos was done using the 488 nm laser on a Marianas Real Time Confocal SDC Workstation spinning disc confocal microscope (Inteligent Imaging Innovations) employing a CSU-X1 Yokogawa scan head and a modified Cascade QuantEM 512SC EMCCD camera. Objective lenses was a Zeiss 100x NA 1.46 oil immersion. Images were acquired using the SlideBook 5.0 software (Inteligent Imaging Innovations) at a rate of ~100 ms/frame.

DIC images from a representative time-lapse sequence of nud-2(ok949) nuclear migration in hyp7 precursors. Cellular boundaries appear as depressions in the hypodermis and nuclei are exclusions of cytoplasmic granules. Embryos were dissected from gravid hermaphrodites and mounted on a 2% agar pad in M9 solution. Time-lapse microscopy of nuclear migration was observed using DIC optics and a 63x PLAN APO 1.40 NA objective on a Leica DM 6000 compound microscope. Images were acquired using a Leica DC350 FX camera and the LAS AF software (Leica) at a rate of 15 sec/frame.

DIC images from a representative time-lapse sequence of unc-83(e1408) nuclear migration in hyp7 precursors. Cellular boundaries appear as depressions in the hypodermis and nuclei are exclusions of cytoplasmic granules. Embryos were dissected from gravid hermaphrodites and mounted on a 2% agar pad in M9 solution. Time-lapse microscopy of nuclear migration was observed using DIC optics and a 63x PLAN APO 1.40 NA objective on a Leica DM 6000 compound microscope. Images were acquired using a Leica DC350 FX camera and the LAS AF software (Leica) at a rate of 15 sec/frame.

DIC images from a representative time-lapse sequence of nuclear migration in hyp7 precursors. Cellular boundaries appear as depressions in the hypodermis and nuclei are exclusions of cytoplasmic granules. Nucleus that rolls is in about the center of the embryo at frame 270 when it begins rolling. Embryos were dissected from gravid hermaphrodites and mounted on a 2% agar pad in M9 solution. Time-lapse microscopy of nuclear migration was observed using DIC optics and a 63x PLAN APO 1.40 NA objective on a Leica DM 6000 compound microscope. Images were acquired using a Leica DC350 FX camera and the LAS AF software (Leica) at a rate of 15 sec/frame.

Gamma-tubulin localizes at cell-cell boundaries. Immunostaining of anti-γ-tubulin in fixed wild-type embryos, costained with DAPI to visualize nuclei. For immunofluorescence, late embryos were extruded from slightly starved hermaphrodites, permeabilized by the freeze-crack method, fixed for 10 minutes in −20 °C methanol, and blocked in PBST (phosphate-buffered saline+0.1% Triton X-100) +5% dry milk. Rabbit polyclonal antibody T5192 was used against γ-tubulin at a 1/200 dilution (Sigma) .

DIC images from a representative time-lapse sequence of wild-type nuclear migration in hyp7 precursors. Cellular boundaries appear as depressions in the hypodermis and nuclei are exclusions of cytoplasmic granules. Embryos were dissected from gravid hermaphrodites and mounted on a 2% agar pad in M9 solution. Time-lapse microscopy of nuclear migration was observed using DIC optics and a 63x PLAN APO 1.40 NA objective on a Leica DM 6000 compound microscope. Images were acquired using a Leica DC350 FX camera and the LAS AF software (Leica) at a rate of 15 sec/frame.

bioimagesImage 16591 (Figure 7 - B)Figure 5 - AFigure 1 - AImage 15541 (Figure 1 - B)Figure 7 - BFigure 5 - BFigure 1 - BFigure 4 - AFigure 5Figure 4Figure 7Image 15542 (Figure 5 - A)Figure 9Figure 9 - NoneImage 15543 (Figure 5 - B)Image 16545 (Figure 9 - None)Figure 1Image 15544 (Figure 4 - A)Image 15452 (Figure 1 - A)Daniel A. StarrHeidi N. FridolfssonfalseKinesin-1 and dynein at the nuclear envelope mediate the bidirectional migrations of nuclei2010-10-04T11:20:24Z2018-11-29T11:20:24Z2018-11-29T11:20:24ZS-JCBD-20100411810.1083/jcb.201004118