Cells were stably transfected with GFP-linked nuclear localization
(NLS-GFP) and cultured in Dulbecco’s modified Eagle’s medium
(DMEM) supplemented with 10% fetal bovine serum (FBS), 1%
penicillin/streptomycin, and 0.5 mg/ml G418. Tissue culture plates were coated
with collagen I and 50,000 cells were added to the center of the well and
allowed to adhere for 24 hours. Cells were maintained at 37C and 5%
CO2 as images were recorded every three minutes for 54 hours
using phase microscopy and confocal fluorescence using the 488 nm line of an
argon laser on a Leica DMI6000 SP5 microscope with a stage top incubator. Images
were processed to find centroids of each nuclei, which were used as seeds for a
Voronoi tessellation in order to create a polygonal tiling of the cell layer.
These polygons were then used to obtain cell aspect ratio using an in-house
custom algorithm.
(NLS-GFP) and cultured in Dulbecco’s modified Eagle’s medium
(DMEM) supplemented with 10% fetal bovine serum (FBS), 1%
penicillin/streptomycin, and 0.5 mg/ml G418. Tissue culture plates were coated
with collagen I and 50,000 cells were added to the center of the well and
allowed to adhere for 24 hours. Cells were maintained at 37C and 5%
CO2 as images were recorded every three minutes for 54 hours
using phase microscopy and confocal fluorescence using the 488 nm line of an
argon laser on a Leica DMI6000 SP5 microscope with a stage top incubator. Images
were processed to find centroids of each nuclei, which were used as seeds for a
Voronoi tessellation in order to create a polygonal tiling of the cell layer.
These polygons were then used to obtain cell aspect ratio using an in-house
custom algorithm.