Abstract
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.
Original language | English (US) |
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Pages (from-to) | 12403-12414 |
Number of pages | 12 |
Journal | Optics Express |
Volume | 16 |
Issue number | 16 |
DOIs | |
State | Published - Aug 4 2008 |
ASJC Scopus subject areas
- Atomic and Molecular Physics, and Optics