在Cyris®分析平台上的体外线粒体解偶联分析和洗脱

IN-VITRO MITOCHONDRIAL UNCOUPLER ASSAY PLUS

WASHOUT ON THE CYRIS® ANALYSIS PLATFORM

作者：Christina Heichler & Dr. Peter Wolf（2021）

Abstract：

The following exemplary protocol describes the preparation, run, and results of an in-vitro assay to investigate the reactions of L929 mouse fibroblasts to treatment and washout of the mitochondrial chain uncoupler carbonyl cyanide-ptrifluoromethoxyphenylhydrazone (FCCP). The CYRIS® platform is able to perform this automated assay with a deeper insight into the substance mode of action due to its ability to monitor the oxygen consumption rate (OCR), extracellular acidification rate (ECAR)and microscopic images of the cells label-free in parallel and in real time. The cells are measured for 12 hours without treatment for baseline, 12 hours under treatment with different FCCP concentrations (0 nM, 63 nM, 125 nM, 250 nM and 500 nM) and 10 hours after treatment for washout. The results show a dose- and time-dependent reaction of the cells to FCCP. Correlations between different cell parameters are clearly visible. It is possible and may be necessary to adapt the protocol to other cell lines, cell numbers, and culture times. It is recommended to read the entire protocol once complete before.

文章摘要：

以下案例描述了体外测定的制备、运行和结果，以研究 L929 小鼠成纤维细胞对线粒体链解偶联剂羰基氰化物-ptrifluoromethoxyphenylhydrazone (FCCP) 的处理和冲洗的反应。

CYRIS® 平台能够监测耗氧率 (OCR)、细胞外酸化率 (ECAR) 和无标记细胞的显微图像，因此能够更深入地了解物质作用模式来执行这种自动化检测和实时数据收集。测量未处理的细胞 12 小时作为基础数据，在不同 FCCP 浓度（0 nM、63 nM、125 nM、250 nM 和 500 nM）处理下 12 小时和处理后 10 小时进行洗脱。

结果显示细胞对 FCCP 的剂量和时间依赖性反应。不同细胞参数之间的相关性清晰可见。有可能并且可能有必要使案例应用于其他细胞系、细胞数量和培养时间。

建议在操作之前阅读整个案例。

点击链接即可查看和下载文章：https://www.cyris.bio/wp-content/uploads/2022/12/20210511-Application-Note-L929-FCCP_my.pdf

文章题目、关键词与摘要译文仅用于参考。