Health and Biomedical Innovation, University of South Australia, Adelaide SA, Australia.
Quality Use of Medicines and Pharmacy Research Centre, UniSA Clinical and Health Sciences, University of South Australia, Adelaide SA, Australia.
Mutagenesis. 2020 Sep 12;35(4):311-318. doi: 10.1093/mutage/geaa013.
The cytokinesis-block micronucleus cytome (CBMNcyt) assay is a comprehensive method to measure DNA damage, cytostasis and cytotoxicity caused by nutritional, radiation and chemical factors. A slide imaging technique has been identified as a new method to assist with the visual scoring of cells for the CBMNcyt assay. A NanoZoomer S60 Digital Pathology slide scanner was used to view WIL2-NS cells treated with hydrogen peroxide (H2O2) and measure CBMNcyt assay biomarkers using a high-definition desktop computer screen. The H2O2-treated WIL2-NS cells were also scored visually using a standard light microscope, and the two visual scoring methods were compared. Good agreement was found between the scoring methods for all DNA damage indices (micronuclei, nucleoplasmic bridges and nuclear buds) and nuclear division index with correlation R values ranging from 0.438 to 0.789, P < 0.05. Apoptotic and necrotic cell frequency was lower for the NanoZoomer scoring method, but necrotic frequency correlated well with the direct visual microscope method (R = 0.703, P < 0.0001). Considerable advantages of the NanoZoomer scoring method compared to direct visual microscopy includes reduced scoring time, improved ergonomics and a reduction in scorer fatigue. This study indicates that a digital slide scanning and viewing technique may assist with visual scoring for the CBMNcyt assay and provides similar results to conventional direct visual scoring.
细胞胞质分裂阻断微核细胞胞质(CBMNcyt)分析是一种综合的方法,用于测量营养、辐射和化学因素引起的 DNA 损伤、细胞停滞和细胞毒性。幻灯片成像技术已被确定为一种新的方法,可用于协助对 CBMNcyt 分析中的细胞进行视觉评分。NanoZoomer S60 数字病理学幻灯片扫描仪用于观察用过氧化氢(H2O2)处理的 WIL2-NS 细胞,并使用高清台式计算机屏幕测量 CBMNcyt 分析生物标志物。还用标准显微镜对 H2O2 处理的 WIL2-NS 细胞进行了目视评分,并比较了两种目视评分方法。对于所有 DNA 损伤指数(微核、核质桥和核芽)和核分裂指数,两种评分方法之间的评分结果具有良好的一致性,相关 R 值范围为 0.438 至 0.789,P<0.05。NanoZoomer 评分方法的细胞凋亡和坏死频率较低,但坏死频率与直接显微镜方法相关性良好(R=0.703,P<0.0001)。与直接目视显微镜相比,NanoZoomer 评分方法具有许多优势,包括评分时间缩短、人体工程学改善和评分员疲劳度降低。本研究表明,数字幻灯片扫描和查看技术可协助 CBMNcyt 分析的目视评分,并提供与传统直接目视评分相似的结果。