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首页> 外文期刊>Scientific reports. >Automated Segmentation of Fluorescence Microscopy Images for 3D Cell Detection in human-derived Cardiospheres
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Automated Segmentation of Fluorescence Microscopy Images for 3D Cell Detection in human-derived Cardiospheres

机译：荧光显微镜图像的自动分割，用于人类源性心球中的3D细胞检测

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The 'cardiosphere' is a 3D cluster of cardiac progenitor cells recapitulating a stem cell niche-like microenvironment with a potential for disease and regeneration modelling of the failing human myocardium. In this multicellular 3D context, it is extremely important to decrypt the spatial distribution of cell markers for dissecting the evolution of cellular phenotypes by direct quantification of fluorescent signals in confocal microscopy. In this study, we present a fully automated method, named CARE ('CARdiosphere Evaluation'), for the segmentation of membranes and cell nuclei in human-derived cardiospheres. The proposed method is tested on twenty 3D-stacks of cardiospheres, for a total of 1160 images. Automatic results are compared with manual annotations and two open-source software designed for fluorescence microscopy. CARE performance was excellent in cardiospheres membrane segmentation and, in cell nuclei detection, the algorithm achieved the same performance as two expert operators. To the best of our knowledge, CARE is the first fully automated algorithm for segmentation inside in vitro 3D cell spheroids, including cardiospheres. The proposed approach will provide, in the future, automated quantitative analysis of markers distribution within the cardiac niche-like environment, enabling predictive associations between cell mechanical stresses and dynamic phenotypic changes.
机译：“心脏层”是心脏祖细胞的3D簇，概括了干细胞小生境样的微环境，具有对衰竭的人类心肌进行疾病和再生建模的潜力。在这种多细胞3D环境中，通过在共聚焦显微镜中直接定量荧光信号来解密细胞标志物的空间分布，以剖析细胞表型的演变，这一点极为重要。在这项研究中，我们提出了一种名为CARE（“ CARdiosphere评估”）的全自动方法，用于分割人源性心球中的膜和细胞核。该方法在20个心球3D堆栈上进行了测试，总共获得1160张图像。将自动结果与手动注释和两个专为荧光显微镜设计的开源软件进行比较。 CARE性能在心球膜分割方面非常出色，在细胞核检测中，该算法的性能与两名专家操作员相同。据我们所知，CARE是第一个用于体外3D细胞球体（包括心球）内部分割的全自动算法。拟议的方法将在将来提供对心脏利基样环境内标记分布的自动定量分析，从而实现细胞机械应力与动态表型变化之间的预测关联。

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