Automated Detection and Analysis of Ca2+ Sparks in x-y Image Stacks Using a Thresholding Algorithm Implemented within the Open-Source Image Analysis Platform ImageJ
Automated Detection and Analysis of Ca2+ Sparks in x-y Image Stacks Using a Thresholding Algorithm Implemented within the Open-Source Image Analysis Platform ImageJ
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DOI：
10.1016/j.bpj.2013.12.040
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发表时间：
2014-02-04
影响因子：
3.4
通讯作者：
Steele, Derek S.
中科院分区：
文献类型：
--
作者：
Steele, Elliot M.;Steele, Derek S.
以前的研究已经广泛地使用钙火花的分析来研究心肌细胞的正常和病理性的钙调节。这些研究中的绝大多数使用了行扫描共聚焦成像。在一定程度上，这是因为用于自动检测行扫描图像中的钙火花的开源软件的开发大大简化了数据分析。行扫描成像的一个缺点是数据是从单行像素收集的，仅代表细胞的一小部分，在许多情况下x-y共焦成像更可取。然而，用于二维x-y图像堆栈的钙离子火花分析软件的有限可用性为其更广泛的应用提供了障碍。这项研究描述了能够自动检测和分析x-y图像堆栈中的钙火花的软件的开发和特性，该软件作为开源图像分析平台ImageJ中的一个插件实现。该程序包括能够精确识别共聚焦荧光图像中的细胞的方法，补偿背景荧光的变化，以及允许基于空间特征排除事件的选项。
Previous studies have used analysis of Ca2+ sparks extensively to investigate both normal and pathological Ca2+ regulation in cardiac myocytes. The great majority of these studies used line-scan confocal imaging. In part, this is because the development of open-source software for automatic detection of Ca2+ sparks in line-scan images has greatly simplified data analysis. A disadvantage of line-scan imaging is that data are collected from a single row of pixels, representing only a small fraction of the cell, and in many instances x-y confocal imaging is preferable. However, the limited availability of software for Ca2+ spark analysis in two-dimensional x-y image stacks presents an obstacle to its wider application. This study describes the development and characterization of software to enable automatic detection and analysis of Ca2+ sparks within x-y image stacks, implemented as a plugin within the open-source image analysis platform ImageJ. The program includes methods to enable precise identification of cells within confocal fluorescence images, compensation for changes in background fluorescence, and options that allow exclusion of events based on spatial characteristics.