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通过数字全息显微镜进行的无标记细胞毒性筛选测定。

Label-free cytotoxicity screening assay by digital holographic microscopy.

作者信息

Kühn Jonas, Shaffer Etienne, Mena Julien, Breton Billy, Parent Jérôme, Rappaz Benjamin, Chambon Marc, Emery Yves, Magistretti Pierre, Depeursinge Christian, Marquet Pierre, Turcatti Gerardo

机构信息

Biomolecular Screening Facility, Swiss Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.

出版信息

Assay Drug Dev Technol. 2013 Mar;11(2):101-7. doi: 10.1089/adt.2012.476. Epub 2012 Oct 12.

Abstract

We introduce a label-free technology based on digital holographic microscopy (DHM) with applicability for screening by imaging, and we demonstrate its capability for cytotoxicity assessment using mammalian living cells. For this first high content screening compatible application, we automatized a digital holographic microscope for image acquisition of cells using commercially available 96-well plates. Data generated through both label-free DHM imaging and fluorescence-based methods were in good agreement for cell viability identification and a Z'-factor close to 0.9 was determined, validating the robustness of DHM assay for phenotypic screening. Further, an excellent correlation was obtained between experimental cytotoxicity dose-response curves and known IC50 values for different toxic compounds. For comparable results, DHM has the major advantages of being label free and close to an order of magnitude faster than automated standard fluorescence microscopy.

摘要

我们介绍了一种基于数字全息显微镜(DHM)的无标记技术,该技术适用于通过成像进行筛选,并展示了其使用哺乳动物活细胞进行细胞毒性评估的能力。对于首个与高内涵筛选兼容的应用,我们将一台数字全息显微镜自动化,以便使用市售的96孔板对细胞进行图像采集。通过无标记DHM成像和基于荧光的方法生成的数据在细胞活力鉴定方面高度一致,并且确定了接近0.9的Z'因子,验证了DHM分析用于表型筛选的稳健性。此外,不同毒性化合物的实验细胞毒性剂量反应曲线与已知IC50值之间获得了极好的相关性。为了获得可比的结果,DHM具有无标记的主要优点,并且比自动化标准荧光显微镜快近一个数量级。

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