Description

A confocal Z-stack from a transgenic Drosophila expressing the kinesin-5 motor KLP61F-GFP embryo during prometaphase phase 1. KOP61-F is required to prevent prometaphase spindle collapse driven by the kinesin-14 Ncd. This image is original data contributing to Fig. 1 "Prometaphase spindle dynamics in Drosophila embryos: steady-state and elongation phase" 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 GFP-KLP61F were collected at 25°C for 1 h, matured for 40 min, dechorionated, placed on heptane glue and covered with halocarbon oil. 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). For these experiments, z-stacks were acquired with an Orca II CCD camera (Hamamatsu Photonics) at intervals of 0.1-0.2 µm, using exposure times of 300-600ms. For further information see J. Cell Biol. 188(1):49-68.