Selection and Tuning of a Fast and Simple Phase-Contrast Microscopy Image Segmentation Algorithm for Measuring Myoblast Growth Kinetics in an Automated Manner
Selection and Tuning of a Fast and Simple Phase-Contrast Microscopy Image Segmentation Algorithm for Measuring Myoblast Growth Kinetics in an Automated Manner
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DOI：
10.1017/s143192761300161x
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发表时间：
2013-08-01
影响因子：
2.8
通讯作者：
Duchesne, Carl
中科院分区：
文献类型：
--
作者：
Juneau, Pierre-Marc;Garnier, Alain;Duchesne, Carl
在宽视场长期活细胞成像和高通量筛选应用中获取和处理相衬显微镜图像仍然是一个挑战，因为所使用的方法和算法必须快速，易于使用和调整，并且尽可能少地侵入。在本文中，我们开发了一个简单而快速的算法来计算相衬显微镜图像中的细胞覆盖表面（汇合度）。该分割算法是基于一个指定大小的范围过滤器，最小范围阈值，和最小目标尺寸阈值。这些参数进行了调整，以最大限度地提高F-测量功能的校准集的200手分割图像，其性能与文献中提出的其他算法进行了比较。对来自37个不同条件下的成肌细胞培养物的一组一百万张图像进行处理，以获得它们随时间变化的细胞覆盖表面。数据被用来拟合指数和逻辑模型，分析显示动力学参数和传代次数之间的线性关系，并突出了培养基质量对细胞生长动力学的影响。该算法可用于实时监测细胞培养和高通量筛选实验后，适当的调整。
Acquiring and processing phase-contrast microscopy images in wide-field long-term live-cell imaging and high-throughput screening applications is still a challenge as the methodology and algorithms used must be fast, simple to use and tune, and as minimally intrusive as possible. In this paper, we developed a simple and fast algorithm to compute the cell-covered surface (degree of confluence) in phase-contrast microscopy images. This segmentation algorithm is based on a range filter of a specified size, aminimum range threshold, and aminimum object size threshold. These parameters were adjusted in order to maximize the F-measure function on a calibration set of 200 hand-segmented images, and its performance was compared with other algorithms proposed in the literature. A set of one million images from 37 myoblast cell cultures under different conditions were processed to obtain their cell-covered surface against time. The data were used to fit exponential and logistic models, and the analysis showed a linear relationship between the kinetic parameters and passage number and highlighted the effect of culture medium quality on cell growth kinetics. This algorithm could be used for real-time monitoring of cell cultures and for high-throughput screening experiments upon adequate tuning.