Three dimensional data set acquired using a fast z-scan along the apical-to-basilar axis of a polarized epithelial cell in vitro to capture GFP-EB1 comets assosiated with the plus-end of actively polymerizing microtubules (green), and the microtubule anchoring factor RFP-LL5α (red). The apical region of the cell is at the bottom of the Z-axis, and basilar region is at the top. This is the original data file from Fig. 8 C from J Cell Biol (2010) 189 (5):901-917.
MCF-10Aeco cells expressing EB1-GFP and RFP-LL5 were cocultured with parental non-fluorescent cells on glass-bottomed dishes (fluorescent:non-fluorescent cells=1:10). Z-scanning images from the apical (bottom of the Z-axis here) to basal planes (top of the Z axis) of the cell were collected using a Revolution XD system driven by IQ software (Andor Technology) equipped with an IX81 inverted microscope with a UPlanSApo 100× NA 1.40 oil immersion objective (Olympus), an electron-multiplying charge-coupled device cam- era (iXon DU-888; Andor Technology), a confocal spinning disk (CSU22; Yokogawa), a piezo-Z stage, a laser combiner with an acousto-optic tun- able filter and a CO2 incubator for live cell culture.
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