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较小规模,相同的影响:在中等吞吐量实验室中复制高吞吐量表型分析,用于化学风险评估

Eunnara Cho¹, Stephen D Baird², Kristin M Eccles³

¹Environmental Health Science and Research Bureau, Healthy Environments and Consumer Safety Branch (HECSB) Health Canada, Ottawa, ON, K1A 0K9, Canada.

Archives of toxicology

|August 28, 2025

概括

细胞绘制是一种高通量表型分析 (HTPP) 的方法,适用于96孔板,对毒性评估显示一致的结果. 这种调整增加了各种实验室中毒性测试的可用性.

科学领域:

细胞成像和分析
毒理学和危险评估
高含量查

背景情况:

使用光染料和高含量成像进行高通量表型分析 (HTPP) 进行毒性评估.
目前的HTPP协议通常优化为384井板,限制低通量实验室的可访问性.
监管机构使用HTPP进行化学危险选.

研究的目的:

将已建立的细胞涂料协议从384孔调整为96孔板.
评估以96孔格式生成的毒性数据的一致性和可比性.
评估细胞绘画在不同实验室格式中的适应性.

主要方法:

用12个参考化合物在96个井板中培养和处理U-2 OS细胞.
细胞被染色为五个细胞区 (Golgi,ER,核酸,细胞骨,线粒体) 和成像.
用主要成分分析和Mahalanobis距离来计算基准度 (BMC) 来提取,规范化和分析形态特征.

主要成果:

经过调整的96孔板协议显示了实验室内部的一致性,大多数BMC的差异不到一个数量级.
在96孔格式中获得的BMC值与12种化合物中的10种 BMC值相比较.
观察到细胞播种密度和Mahalanobis距离之间的反向关系,表明实验因素的潜在影响.

结论:

细胞涂料可以适应96个井板格式,从而提高其对毒性测试的可访问性.
采用96孔细胞涂料的方法,可提供与已知方法一致且可比的结果.
这种调整支持细胞涂料的验证,作为毒性测试的补充新方法.

关键词:

细胞绘画高内容成像高通量表型分析毒性评估

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