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Quantification of three-dimensional dynamics of intercellular geometry under mechanical loading using a weighted directional adaptive-threshold method

机译:使用加权方向自适应阈值方法量化机械载荷下细胞间几何结构的三维动力学

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摘要

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.
机译:由于成像系统的机械限制以及时间分辨率和图像质量之间的固有权衡,在快速的时间尺度上捕获和量化三维细胞几何结构中的动态变化是一个挑战。我们将定制的高速双光子显微镜方法与一种新颖的图像分割方法(加权方向自适应阈值(WDAT))相结合,以量化压缩压力下以前无法访问的时间尺度下细胞胞间空间的大小。高速双光子显微镜的适应性解决了捕获短时间尺度上发生的事件的需求,而WDAT方法被开发用于处理应用于该系统的基于标准强度的分析方法的伪像。我们的新方法通过对气道上皮细胞压缩负荷伴随的三维细胞和细胞外变形的增强时间分析得到证明。

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