Border Samuel, Rosenberg Avi, Zee Jarcy, Levenson Richard, Jen Kuang-Yu, Sarder Pinaki, Fereidouni Farzad
University of Florida at Gainesville, J. Crayton Pruitt Family Department of Biomedical Engineering.
Johns Hopkins Medicine, Department of Pathology.
Proc SPIE Int Soc Opt Eng. 2023 Feb;12471. doi: 10.1117/12.2654651. Epub 2023 Apr 6.
Accurate quantification of renal fibrosis has profound importance in the assessment of chronic kidney disease (CKD). Visual analysis of a biopsy stained with trichrome under the microscope by a pathologist is the gold standard for evaluation of fibrosis. Trichrome helps to highlight collagen and ultimately interstitial fibrosis. However, trichrome stains are not always reproducible, can underestimate collagen content and are not sensitive to subtle fibrotic patterns. Using the Dual-mode emission and transmission (DUET) microscopy approach, it is possible to capture both brightfield and fluorescence images from the same area of a tissue stained with hematoxylin and eosin (H&E) enabling reproducible extraction of collagen with high sensitivity and specificity. Manual extraction of spectrally overlapping collagen signals from tubular epithelial cells and red blood cells is still an intensive task. We employed a UNet++ architecture for pixel-level segmentation and quantification of collagen using 760 whole slide image (WSI) patches from six cases of varying stages of fibrosis. Our trained model (Deep-DUET) used the supervised extracted collagen mask as ground truth and was able to predict the extent of collagen signal with a MSE of 0.05 in a holdout testing set while achieving an average AUC of 0.94 for predicting regions of collagen deposits. Expanding this work to the level of the WSI can greatly improve the ability of pathologists and machine learning (ML) tools to quantify the extent of renal fibrosis reproducibly and reliably.
肾纤维化的准确量化在慢性肾脏病(CKD)评估中具有深远意义。病理学家在显微镜下对经三色染色的活检组织进行视觉分析是评估纤维化的金标准。三色染色有助于突出胶原蛋白并最终显示间质纤维化。然而,三色染色并不总是可重复的,可能会低估胶原蛋白含量,并且对细微的纤维化模式不敏感。使用双模式发射和透射(DUET)显微镜方法,可以从用苏木精和伊红(H&E)染色的组织的同一区域捕获明场和荧光图像,从而以高灵敏度和特异性可重复地提取胶原蛋白。从肾小管上皮细胞和红细胞中手动提取光谱重叠的胶原蛋白信号仍然是一项繁重的任务。我们采用UNet++架构对来自六个不同纤维化阶段病例的760个全切片图像(WSI)补丁进行胶原蛋白的像素级分割和量化。我们训练的模型(Deep-DUET)使用监督提取的胶原蛋白掩码作为基本事实,在保留测试集中能够以0.05的均方误差预测胶原蛋白信号的程度,同时在预测胶原蛋白沉积区域时平均AUC达到0.94。将这项工作扩展到WSI水平可以大大提高病理学家和机器学习(ML)工具可重复且可靠地量化肾纤维化程度的能力。