PIV as a Method for Quantifying Root Cell Growth and Particle Displacement in Confocal Images
PIV as a Method for Quantifying Root Cell Growth and Particle Displacement in Confocal Images
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DOI：
10.1002/jemt.20749
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发表时间：
2010-01-01
影响因子：
2.5
通讯作者：
Valentine, Tracy A.
中科院分区：
文献类型：
--
作者：
Bengough, A. Glyn;Hans, Joachim;Valentine, Tracy A.
粒子图像测速技术（PIV）量化连续图像之间像素块的位移。我们评估PIV作为一种工具，显微镜通过测量位移的细胞和周围的颗粒介质在共聚焦激光扫描显微镜图像标记的细胞膜靶向绿色荧光蛋白的拟南芥根。出色的准确性（例如，位移标准差0.83）。这些失真图像的均方根误差为0.4-1.1像素，在更高的放大倍数下增加。细胞生长和根际变形以良好的时间（例如，1-最小间隔）和空间分辨率，位于可识别细胞特征上的PIV斑块可以更成功地跟踪。选择合适的GFP标记物对减少细胞内运动引起的小尺度生物噪声具有重要意义。PIV的根生长在硬2%与0.7%琼脂显示模式的细胞扩张与其他物种的物理阻碍根一致。根在玻璃ballotini经历了快速的变化，在生长方向上的时间尺度分钟，与本地化的ballotini。通过跟踪细胞顶点，我们自动监测细胞的长度，宽度和面积每分钟为0.5小时的细胞在不同的发展阶段。总之，PIV成功地测量了活根细胞和外部颗粒介质图像中的位移，揭示了显微镜工作者使用的巨大潜力。Microsc. Res. Tech. 73：27-36，2010. (C)2009威利-利斯公司
Particle image velocimetry (PIV) quantifies displacement of patches of pixels between successive images. We evaluated PIV as a tool for microscopists by measuring displacements of cells and of a surrounding granular medium in confocal laser scanning microscopy images of Arabidopsis thaliana roots labeled with cell-membrane targeted green fluorescent protein. Excellent accuracy (e.g., displacement standard deviation 0.83). Root mean squared error for these distorted images was 0.4-1.1 pixels, increasing at higher magnification factors. Cell growth and rhizosphere deformation were tracked with good temporal (e.g., 1-min interval) and spatial resolution, with PIV patches located on recognizable cell features being tracked more successfully. Appropriate choice of GFP-label was important to decrease small-scale biological noise due to intracellular motion. PIV of roots grown in stiff 2% versus 0.7% agar showed patterns of cell expansion consistent with physically impeded roots of other species. Roots in glass ballotini underwent rapid changes in growth direction on a timescale of minutes, associated with localized arching of ballotini. By tracking cell vertices, we monitored automatically cell length, width, and area every minute for 0.5 h for cells in different stages of development. In conclusion, PIV measured displacements successfully in images of living root cells and the external granular medium, revealing much potential for use by microscopists. Microsc. Res. Tech. 73:27-36, 2010. (C) 2009 Wiley-Liss, Inc.