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阿拉玛蓝检测优化，以尽量减少药物干扰和检测间的活力差异。

Alamar Blue assay optimization to minimize drug interference and inter assay viability.

作者信息

Dinh Mina N, Hitomi Masahiro, Al-Turaihi Zahraa A, Scott Jacob G

机构信息

Department of Translational Hematology & Oncology Research, Cleveland Clinic, Cleveland, OH, United States of America.

Department of Radiation Oncology, Cleveland Clinic, Cleveland, OH, United States of America.

出版信息

MethodsX. 2024 Oct 28;13:103024. doi: 10.1016/j.mex.2024.103024. eCollection 2024 Dec.

DOI:10.1016/j.mex.2024.103024
PMID:39553738
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11566963/
Abstract

Alamar Blue (AB) has become an increasingly popular reagent of choice for cell viability assays. We chose AB over other reagents such as MTT and Cell-Titer Glo due to its cost effectiveness and its ability to be a nondestructive assay. While analyzing the effect of osimertinib, an EGFR inhibitor on the non-small cell lung cancer cell line, PC-9, we noticed unexpected right-shifts of dose response curves as compared to the curves obtained by Cell Titer Glo assay. Here, we describe our modified AB assay method to avoid that right shift in dose response curves.•Removal of the drug containing medium prior to AB addition eliminated the falsely increased readings, giving comparable dose response curves as determined by Cell Titer Glo assay.•Plate-to-plate variability can be minimized by adding an appropriate concentration of a common fluorescence calibration standard to the assay plates to calibrate fluorimeter sensitivity.•Alamar Blue can be used as a continuous longitudinal assay to monitor cell growth or recovery from drug toxicity over time.

摘要

alamar蓝（AB）已成为细胞活力测定中越来越受欢迎的首选试剂。由于其成本效益和无损检测能力，我们选择AB而非其他试剂，如MTT和Cell-Titer Glo。在分析表皮生长因子受体（EGFR）抑制剂奥希替尼对非小细胞肺癌细胞系PC-9的影响时，我们注意到与Cell Titer Glo检测获得的曲线相比，剂量反应曲线出现了意外的右移。在此，我们描述了我们改进的AB检测方法，以避免剂量反应曲线的右移。

  • 在添加AB之前去除含药培养基可消除错误增加的读数，得到与Cell Titer Glo检测相当的剂量反应曲线。

  • 通过向检测板中添加适当浓度的通用荧光校准标准物以校准荧光计灵敏度，可将板间差异降至最低。

  • alamar蓝可用于连续纵向检测，以监测细胞生长或随时间从药物毒性中恢复的情况。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20c9/11566963/6fd80f91b8c6/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20c9/11566963/f93fd9643969/ga1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20c9/11566963/55a86f7862fb/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20c9/11566963/08705cf34327/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20c9/11566963/45cf5c059ff4/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20c9/11566963/6fd80f91b8c6/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20c9/11566963/f93fd9643969/ga1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20c9/11566963/55a86f7862fb/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20c9/11566963/08705cf34327/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20c9/11566963/45cf5c059ff4/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20c9/11566963/6fd80f91b8c6/gr4.jpg

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