Spinning disk confocal time-lapse imaging of the first mitosis in a C. elegans embryo expressing GFP-CeCENP-A in which KNL-2 has been depleted by RNAi. GFP-CeCENP-A was imaged (strain OD101). Diffuse nuclear GFP signal is evident during prophase. However, unlike the control (CIL 28780), no discrete GFP foci were seen at any point. The video is ∼12 min long, and the frame is ∼30 µm top to bottom. DIC is shown on the left, and GFP is shown on the right.
Images were acquired via a spinning disk confocal microscope (CSU10; McBain Instruments) mounted on an inverted microscope (TE2000e; Nikon). Strain TH32 coexpressing GFP-histone H2b and GFP–γ-tubulin was imaged using a 60× 1.4 NA plan Apo objective with 1.5× auxiliary magnification and a cooled CCD camera (Orca ER; Hamamatsu) binning 2 × 2. Strain OD31 expressing GFP–KNL-2 was imaged in the same manner without the 1.5× auxiliary magnification.
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