Effect of Cell Shape and Dimensionality on Spindle Orientation and Mitotic Timing

Brief description

Time lapse microscopy images of HeLa cells dividing in different shaped microwells.
This central aim of this project was to determine the effect of 3D cell shape on mitosis. To achieve this, HeLa cells were cultured in square versus circular microwells and imaged while undergoing mitosis using time lapse microscopy.

For time lapse imaging, cells were monitored in Leibovitz's L-15 medium supplemented with fetal calf serum (10% (v/v)), penicillin (100 U/ml), streptomycin (100 μg/ml) and amphotericin (0.625 μg/ml). Time points, comprised of 10 z sections 1 µm apart, were acquired every 4 minutes for 8 hours with a 63× objective lens (1.4NA DIC oil PlanApo, Olympus) and a camera (CoolSNAP HQ; Roper Scientific) on an imaging system (DeltaVision Core, Applied Precision) fitted with a 37°C environmental chamber. Image stacks were deconvolved and quantified with SoftWorx (Applied Precision, LLC) and mounted in figures using Imaris and Photoshop.

Data and Access: 100 files stored as axiovision time lapse movies.

This research was conducted at ETH Zurich and funded by grants from Swiss National Science Foundation (SNSF; www.snf.ch; grant number CR23I2_125290/1), the Competence Centre for Materials Science and Technoloy (CCMX; www.ccmx.ch), and the National Centers for Competence in Research (NCCR; www.nccr.ch; Nano-P5, project 03). Related Publications: This data has been published in PLoS ONE