Department of Surgery, National Taiwan University Hospital Yun-Lin Branch, Yun-Lin County, Taiwan; Graduate Institute of Clinical Medicine, National Taiwan University College of Medicine, Taipei, Taiwan.
HistoIndex Pte Ltd, Singapore.
Clin Res Hepatol Gastroenterol. 2020 Feb;44(1):12-20. doi: 10.1016/j.clinre.2019.04.003. Epub 2019 May 7.
Quantitative assessments of liver fibrosis using second-harmonic generation/two-photon excited fluorescence microscopy provide greater sensitivity and accuracy than collagen proportionate area while eliminating operator-dependent variation in the staining process. In conjunction with sophisticated image analysis algorithms and feature selection, we might reduce the computation cost in future and narrow down the candidates for further clinical studies.
We sampled a total of 244 liver specimens from patients with hepatitis B viral infections who underwent liver biopsy or liver resection at the National Taiwan University Hospital. The samples were then imaged using a Genesis (HistoIndex Pte. Ltd, Singapore) system, wherein second-harmonic generation microscopy was used to visualize collagen, and two-photon excited fluorescence microscopy was used to visualize other cell structures. We used 100 morphological features extracted from the images to assess correlations with METAVIR fibrosis scores.
Out of 100 quantitative measurements, 76 showed significant correlation with METAVIR scoring, thereby enabling the statistical discrimination of patients in various stages of the disease. These 76 features were also narrowed down by the nonlinear test to 10 candidate measurements, which can be further investigated in detail.
Our experimental results showed that the model with 10 selected features can beat the one with second-harmonic generation only, and performed equivalently well compared the model with 76 features, especially for early-stage discrimination. Features presenting significant correlation were used to fit a single combined index in order to predict pathological staging, thereby making it possible to reveal incremental progress during treatment.
使用二次谐波产生/双光子激发荧光显微镜对肝纤维化进行定量评估，其灵敏度和准确性高于胶原比例面积，同时消除了染色过程中操作人员依赖性的变化。结合复杂的图像分析算法和特征选择，我们可能会降低未来的计算成本，并缩小进一步临床研究的候选者范围。
我们从在台湾大学医院接受肝活检或肝切除术的乙型肝炎病毒感染患者中总共采集了 244 个肝标本。然后使用 Genesis（HistoIndex Pte. Ltd，新加坡）系统对这些样本进行成像，其中二次谐波产生显微镜用于可视化胶原，双光子激发荧光显微镜用于可视化其他细胞结构。我们使用从图像中提取的 100 个形态特征来评估与 METAVIR 纤维化评分的相关性。
在 100 个定量测量中，有 76 个与 METAVIR 评分显著相关，从而能够对不同疾病阶段的患者进行统计区分。这些 76 个特征也通过非线性测试缩小到 10 个候选测量值，可以进一步详细研究。
我们的实验结果表明，具有 10 个选定特征的模型可以击败仅具有二次谐波的模型，并且与具有 76 个特征的模型表现相当，特别是在早期阶段的区分。具有显著相关性的特征用于拟合单个组合指标以预测病理分期，从而有可能揭示治疗过程中的增量进展。
