A confocal 4D-stack from a transgenic Drosophila embryo expressing lamin-GFP (green) and injected with rhodamine-conjugated tubulin (red) and anti-lamin-B antibodies during prometaphase. The microinjection of a mixture of anti-lamin-B mAbs crosslinks the lamin-B envelope into a hyperstable network that fails to disassemble and remains throughout mitosis. This structure impedes spindle elongation. This image is original data contributing to Fig. 6 "Functional perturbation of the lamin-B envelope interferes with spindle length changes and the completion of mitosis" from Civelekoglu-Scholey et al.(2010) Prometaphase spindle maintenance by an antagonistic motor-dependent force balance made robust by a disassembling lamin-B envelope, J. Cell Biol. 188(1):49-68.
Embryos expressing lamin-B-GFP were collected at 25°C for 1 h, matured for 40 min, dechorionated, placed on heptane glue and covered with halocarbon oil. For injections, embryos were dehydrated for 3-6 min, covered with halocarbon oil and injected (as described in Brust-Mascher & Scholey, 2009). For double injections, embryos were allowed to recover for 5–20 min between injections. Images from this image group were acquired with an inverted IX-70 Olympus with Ultra-View spinning disk confocal head (PerkinElmer) and acquired with an oil immersion objective (UPlan-Apochromat 100x N.A. 1.35, or Plan-Apochromat 60x NA 1.4). Time series (at intervals of 3 to 10 sec) z-stacks of planes at 0.5 µm were acquired with an Orca II CCD camera (Hamamatsu Photonics). For further information see J. Cell Biol. 188(1):49-68.
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