An Image Recognition-Based Approach to Actin Cytoskeleton Quantification
An Image Recognition-Based Approach to Actin Cytoskeleton Quantification
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DOI：
10.3390/electronics7120443
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发表时间：
2018-12-01
期刊：
影响因子：
2.9
通讯作者：
Ren, Juan
中科院分区：
文献类型：
--
作者：
Liu, Yi;Mollaeian, Keyvan;Ren, Juan
肌动蛋白细胞骨架的定量对于揭示细胞力的感知和转导机制是至关重要的。虽然荧光成像为观察肌动蛋白细胞骨架的形态提供了一种方便的工具，但由于缺乏准确的肌动蛋白细胞骨架定量方法，对机械转导的动力学仍知之甚少。目前，现有的基于图像的肌动蛋白细胞骨架分析工具要么不能同时定量肌动蛋白细胞骨架的方向和数量，要么量化结果受到分析伪影的影响。在这项研究中，我们提出了一种基于图像识别的肌动蛋白细胞骨架量化方法，该方法通过边缘、直线和亮度检测算法来量化肌动蛋白细胞骨架的方向和数量。通过三个参数来量化肌动蛋白细胞骨架：部分肌动蛋白-细胞骨架偏差(PAD)、总肌动蛋白-细胞骨架偏差(TAD)和平均肌动蛋白-细胞骨架强度(AAI)。首先利用Canny和Sobel边缘检测算子对肌动蛋白细胞骨架图像进行骨架提取，然后利用Hough变换检测出的线条方向对PAD和TAD进行量化，并根据检测到的细胞区域的总和亮度计算AAI。为了验证量化的准确性，建议的伊拉克被应用于六个人工产生的肌动蛋白细胞骨架网状结构模型。量化的PAD和TAD的平均误差均小于1.22度。然后，伊拉克被用来量化用F-肌动蛋白抑制剂(Latrunculin B)处理的NIH/3T3细胞的肌动蛋白细胞骨架。定量结果表明，随着乳链菌素B剂量的增加，局部和整体肌动蛋白细胞骨架结构变得更加无序，肌动蛋白细胞骨架的数量随乳链菌素B剂量的增加而单调减少。
Quantification of the actin cytoskeleton is of prime importance to unveil the cellular force sensing and transduction mechanism. Although fluorescence imaging provides a convenient tool for observing the morphology of the actin cytoskeleton, due to the lack of approaches to accurate actin cytoskeleton quantification, the dynamics of mechanotransduction is still poorly understood. Currently, the existing image-based actin cytoskeleton analysis tools are either incapable of quantifying both the orientation and the quantity of the actin cytoskeleton simultaneously or the quantified results are subject to analysis artifacts. In this study, we propose an image recognition-based actin cytoskeleton quantification (IRAQ) approach, which quantifies both the actin cytoskeleton orientation and quantity by using edge, line, and brightness detection algorithms. The actin cytoskeleton is quantified through three parameters: the partial actin-cytoskeletal deviation (PAD), the total actin-cytoskeletal deviation (TAD), and the average actin-cytoskeletal intensity (AAI). First, Canny and Sobel edge detectors are applied to skeletonize the actin cytoskeleton images, then PAD and TAD are quantified using the line directions detected by Hough transform, and AAI is calculated through the summational brightness over the detected cell area. To verify the quantification accuracy, the proposed IRAQ was applied to six artificially-generated actin cytoskeleton mesh work models. The average error for both the quantified PAD and TAD was less than 1.22 degrees. Then, IRAQ was implemented to quantify the actin cytoskeleton of NIH/3T3 cells treated with an F-actin inhibitor (latrunculin B). The quantification results suggest that the local and total actin-cytoskeletal organization became more disordered with the increase of latrunculin B dosage, and the quantity of the actin cytoskeleton showed a monotonically decreasing relation with latrunculin B dosage.