Design and validation of a tool for neurite tracing and analysis in fluorescence microscopy images
Design and validation of a tool for neurite tracing and analysis in fluorescence microscopy images
复制标题
DOI：
10.1002/cyto.a.20022
复制
发表时间：
2004-04-01
期刊：
影响因子：
3.7
通讯作者：
Unser, M
中科院分区：
文献类型：
--
作者：
Meijering, E;Jacob, M;Unser, M
背景资料：对于神经突生长和分化的分子机制的调查，准确和可重复的分割和定量的神经元过程是一个先决条件。为了促进这项任务，我们开发了一种半自动神经突追踪技术。本文介绍了设计和验证的technology.Methods：该技术进行了比较，完全手动描绘。四名观察员使用这两种方法在培养细胞的20个荧光显微镜图像中重复追踪选定的神经突。准确性和再现性，通过比较高分辨率的参考描记，使用两个错误的措施。劳动密集度是以所需的鼠标点击次数来衡量的。结果的显着性由学生t-检验和方差分析确定。结果：这两种方法略有低估了真实的神经突长度，但差异并不一致显着。与完全手动追踪相比，使用开发的技术与真实神经突中心线的平均偏差小2.6倍。在各自的措施观察员内变异性降低了6.0和23.2倍。观察者间变异性分别降低了2.4倍和8.8倍，劳动强度降低了3.3倍。结论：该技术在测量神经突长度方面具有相似的准确性，显著提高了神经突中心线提取的准确性，显著提高了重复性，降低了劳动强度，可以取代完全手工追踪方法。(C)2004 Wiley-Liss，Inc.
Background: For the investigation of the molecular mechanisms involved in neurite outgrowth and differentiation, accurate and reproducible segmentation and quantification of neuronal processes are a prerequisite. To facilitate this task, we developed a semiautomatic neurite tracing technique. This article describes the design and validation of the technique.Methods: The technique was compared to fully manual delineation. Four observers repeatedly traced selected neurites in 20 fluorescence microscopy images of cells in culture, using both methods. Accuracy and reproducibility were determined by comparing the tracings to high-resolution reference tracings, using two error measures. Labor intensiveness was measured in numbers of mouse clicks required. The significance of the results was determined by a Student t-test and by analysis of variance.Results: Both methods slightly underestimated the true neurite length, but the differences were not unanimously significant. The average deviation from the true neurite centerline was a factor 2.6 smaller with the developed technique compared to fully manual tracing. Intraobserver variability in the respective measures was reduced by a factor 6.0 and 23.2. Interobserver variability was reduced by a factor 2.4 and 8.8, respectively, and labor intensiveness by a factor 3.3.Conclusions: Providing similar accuracy in measuring neurite length, significantly improved accuracy in neurite centerline extraction, and significantly improved reproducibility and reduced labor intensiveness, the developed technique may replace fully manual tracing methods. (C) 2004 Wiley-Liss, Inc.