Aboy-Pardal María C M, Jimenez-Carretero Daniel, Terrés-Domínguez Sara, Pavón Dácil M, Sotodosos-Alonso Laura, Jiménez-Jiménez Víctor, Sánchez-Cabo Fátima, Del Pozo Miguel A
Mechanoadaptation and Caveolae Biology lab, Cell and Developmental Biology Area. Centro Nacional de Investigaciones Cardiovasculares (CNIC), 28029 Madrid, Spain.
Bioinformatics Unit, Centro Nacional de Investigaciones Cardiovasculares (CNIC), 28029 Madrid, Spain.
Comput Struct Biotechnol J. 2022 Dec 5;21:224-237. doi: 10.1016/j.csbj.2022.11.062. eCollection 2023.
Caveolae are nanoscopic and mechanosensitive invaginations of the plasma membrane, essential for adipocyte biology. Transmission electron microscopy (TEM) offers the highest resolution for caveolae visualization, but provides complicated images that are difficult to classify or segment using traditional automated algorithms such as threshold-based methods. As a result, the time-consuming tasks of localization and quantification of caveolae are currently performed manually. We used the Keras library in R to train a convolutional neural network with a total of 36,000 TEM image crops obtained from adipocytes previously annotated manually by an expert. The resulting model can differentiate caveolae from non-caveolae regions with a 97.44% accuracy. The predictions of this model are further processed to obtain caveolae central coordinate detection and cytoplasm boundary delimitation. The model correctly finds negligible caveolae predictions in images from caveolae depleted Cav1 adipocytes. In large reconstructions of adipocyte sections, model and human performances are comparable. We thus provide a new tool for accurate caveolae automated analysis that could speed up and assist in the characterization of the cellular mechanical response.
小窝是质膜的纳米级且对机械敏感的内陷结构,对脂肪细胞生物学至关重要。透射电子显微镜(TEM)为小窝可视化提供了最高分辨率,但所提供的复杂图像难以使用基于阈值的方法等传统自动化算法进行分类或分割。因此,目前小窝定位和定量的耗时任务是手动完成的。我们使用R中的Keras库,用从脂肪细胞中获得的总共36000个TEM图像块训练了一个卷积神经网络,这些图像块此前由一位专家手动标注。所得模型区分小窝和非小窝区域的准确率为97.44%。该模型的预测结果经过进一步处理,以获得小窝中心坐标检测和细胞质边界划定。该模型在来自小窝缺失的Cav1脂肪细胞的图像中正确地发现了可忽略不计的小窝预测。在脂肪细胞切片的大型重建中,模型和人工的表现相当。因此,我们提供了一种用于准确自动分析小窝的新工具,它可以加快并辅助细胞机械反应的表征。