Description

A confocal 4D-stack from a transgenic Drosophila embryo expressing lamin-GFP (green) and injected with rhodamine-conjugated tubulin (red) during prometaphase shows normal elongation of the spindle during the transition during prometaphase II. 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.

Technical Details

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). 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.