Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.
Faculty of Medicine, Albert Ludwigs University of Freiburg, Freiburg, Germany.
Sci Rep. 2021 May 13;11(1):10304. doi: 10.1038/s41598-021-89895-w.
Automated cell classification in cancer biology is a challenging topic in computer vision and machine learning research. Breast cancer is the most common malignancy in women that usually involves phenotypically diverse populations of breast cancer cells and an heterogeneous stroma. In recent years, automated microscopy technologies are allowing the study of live cells over extended periods of time, simplifying the task of compiling large image databases. For instance, there have been several studies oriented towards building machine learning systems capable of automatically classifying images of different cell types (i.e. motor neurons, stem cells). In this work we were interested in classifying breast cancer cells as live or dead, based on a set of automatically retrieved morphological characteristics using image processing techniques. Our hypothesis is that live-dead classification can be performed without any staining and using only bright-field images as input. We tackled this problem using the JIMT-1 breast cancer cell line that grows as an adherent monolayer. First, a vast image set composed by JIMT-1 human breast cancer cells that had been exposed to a chemotherapeutic drug treatment (doxorubicin and paclitaxel) or vehicle control was compiled. Next, several classifiers were trained based on well-known convolutional neural networks (CNN) backbones to perform supervised classification using labels obtained from fluorescence microscopy images associated with each bright-field image. Model performances were evaluated and compared on a large number of bright-field images. The best model reached an AUC = 0.941 for classifying breast cancer cells without treatment. Furthermore, it reached AUC = 0.978 when classifying breast cancer cells under drug treatment. Our results highlight the potential of machine learning and computational image analysis to build new diagnosis tools that benefit the biomedical field by reducing cost, time, and stimulating work reproducibility. More importantly, we analyzed the way our classifiers clusterize bright-field images in the learned high-dimensional embedding and linked these groups to salient visual characteristics in live-dead cell biology observed by trained experts.
癌症生物学中的自动细胞分类是计算机视觉和机器学习研究中的一个具有挑战性的课题。乳腺癌是女性最常见的恶性肿瘤,通常涉及表型多样的乳腺癌细胞群体和异质的基质。近年来,自动化显微镜技术允许对活细胞进行长时间的研究,简化了编译大型图像数据库的任务。例如,已经有几项研究致力于构建能够自动分类不同细胞类型(即运动神经元、干细胞)图像的机器学习系统。在这项工作中,我们有兴趣根据使用图像处理技术自动提取的一组形态特征,对乳腺癌细胞进行活/死分类。我们的假设是,无需任何染色且仅使用明场图像作为输入,即可进行活/死分类。我们使用 JIMT-1 乳腺癌细胞系来解决这个问题,该细胞系以贴壁单层的方式生长。首先,编译了由 JIMT-1 人类乳腺癌细胞组成的大量图像集,这些细胞已经暴露于化疗药物(多柔比星和紫杉醇)或载体对照处理中。接下来,基于著名的卷积神经网络(CNN)骨干,训练了几个分类器,以使用与每个明场图像相关联的荧光显微镜图像的标签进行有监督分类。在大量明场图像上评估和比较模型性能。在未进行治疗的情况下,最佳模型对乳腺癌细胞的分类达到 AUC = 0.941。此外,在对乳腺癌细胞进行药物治疗时,它达到 AUC = 0.978。我们的结果强调了机器学习和计算图像分析的潜力,可以构建新的诊断工具,通过降低成本、时间并激发工作可重复性,使生物医学领域受益。更重要的是,我们分析了我们的分类器在学习的高维嵌入中聚类明场图像的方式,并将这些组与经过训练的专家观察到的活/死细胞生物学中的显著视觉特征联系起来。