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首页> 外文期刊>Physics in medicine and biology. >Automated cell colony counting and analysis using the circular Hough image transform algorithm (CHiTA).
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Automated cell colony counting and analysis using the circular Hough image transform algorithm (CHiTA).

机译：使用循环霍夫图像变换算法（CHiTA）自动进行细胞集落计数和分析。

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摘要

We present an automated cell colony counting method that is flexible, robust and capable of providing more in-depth clonogenic analysis than existing manual and automated approaches. The full form of the Hough transform without approximation has been implemented, for the first time. Improvements in computing speed have facilitated this approach. Colony identification was achieved by pre-processing the raw images of the colonies in situ in the flask, including images of the flask edges, by erosion, dilation and Gaussian smoothing processes. Colony edges were then identified by intensity gradient field discrimination. Our technique eliminates the need for specialized hardware for image capture and enables the use of a standard desktop scanner for distortion-free image acquisition. Additional parameters evaluated included regional colony counts, average colony area, nearest neighbour distances and radial distribution. This spatial and qualitative information extends the utility of the clonogenic assay, allowing analysis of spatially-variant cytotoxic effects. To test the automated system, two flask types and three cell lines with different morphology, cell size and plating density were examined. A novel Monte Carlo method of simulating cell colony images, as well as manual counting, were used to quantify algorithm accuracy. The method was able to identify colonies with unusual morphology, to successfully resolve merged colonies and to correctly count colonies adjacent to flask edges.
机译：我们提出了一种自动化的细胞集落计数方法，该方法灵活，强大并且比现有的手动方法和自动化方法能够提供更深入的克隆形成分析。首次实现了没有近似值的完整Hough变换形式。计算速度的提高促进了这种方法。通过预处理烧瓶中原位菌落的原始图像（包括烧瓶边缘的图像），通过腐蚀，膨胀和高斯平滑处理，可以实现菌落识别。然后通过强度梯度场鉴别来鉴定菌落边缘。我们的技术消除了对用于图像捕获的专用硬件的需求，并使标准台式扫描仪可以用于无失真图像采集。评估的其他参数包括区域菌落计数，平均菌落面积，最近邻距离和径向分布。这种空间和定性信息扩展了克隆形成测定法的用途，从而可以分析空间变异的细胞毒性作用。为了测试自动化系统，检查了两种烧瓶类型和三种具有不同形态，细胞大小和接种密度的细胞系。一种模拟细胞集落图像的新型蒙特卡洛方法以及人工计数被用于量化算法的准确性。该方法能够鉴定出形态异常的菌落，成功解析合并的菌落并正确计数邻近烧瓶边缘的菌落。

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