Quantification of three-dimensional dynamics of intercellular geometry under mechanical loading using a weighted directional adaptive-threshold method.
Capturing and quantifying dynamic changes in three-dimensional cellular geometries on fast time scales is a challenge because of mechanical limitations of imaging systems as well as of the inherent tradeoffs between temporal resolution and image quality. We have combined a custom high-speed two-photon microscopy approach with a novel image segmentation method, the weighted directional adaptive-threshold (WDAT), to quantify the dimensions of intercellular spaces of cells under compressive stress on timescales previously inaccessible. The adaptation of a high-speed two-photon microscope addressed the need to capture events occurring on short timescales, while the WDAT method was developed to address artifacts of standard intensity-based analysis methods when applied to this system. Our novel approach is demonstrated by the enhanced temporal analysis of the three-dimensional cellular and extracellular deformations that accompany compressive loading of airway epithelial cells.
Kojic
Nikola
N
Harvard-MIT Division of Health Sciences and Technology, 77 Mass. Ave., MA 02139, USA.
Huang
Austin
A
Chung
Euiheon
E
Tschumperlin
Daniel
D
So
Peter T
PT
D000465:Algorithms; D002478:Cells, Cultured; D019245:Compressive Strength / Q000502:physiology; D004548:Elasticity; D004847:Epithelial Cells / Q000166:cytology* / Q000502:physiology*; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D018613:Microscopy, Confocal / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D013314:Stress, Mechanical