Rappaz Benjamin, Breton Billy, Shaffer Etienne, Turcatti Gerardo
Biomolecular Screening Facility, Ecole Polytechnique Fédérale de Lausanne (EPFL), Station 15, Lausanne 1015, Switzerland.
Comb Chem High Throughput Screen. 2014 Jan;17(1):80-8. doi: 10.2174/13862073113166660062.
Digital Holographic Microscopy (DHM) is a label-free imaging technique allowing visualization of transparent cells with classical imaging cell culture plates. The quantitative DHM phase contrast image provided is related both to the intracellular refractive index and to cell thickness. DHM is able to distinguish cellular morphological changes on two representative cell lines (HeLa and H9c2) when treated with doxorubicin and chloroquine, two cytotoxic compounds yielding distinct phenotypes. We analyzed parameters linked to cell morphology and to the intracellular content in endpoint measurements and further investigated them with timelapse recording. The results obtained by DHM were compared with other optical label-free microscopy techniques, namely Phase Contrast, Differential Interference Contrast and Transport of Intensity Equation (reconstructed from three bright-field images). For comparative purposes, images were acquired in a common 96-well plate format on the different motorized microscopes. In contrast to the other microscopies assayed, images generated with DHM can be easily quantified using a simple automatized on-the-fly analysis method for discriminating the different phenotypes generated in each cell line. The DHM technology is suitable for the development of robust and unbiased image-based assays.
数字全息显微镜（DHM）是一种无标记成像技术，可通过传统成像细胞培养板实现透明细胞的可视化。所提供的定量DHM相衬图像与细胞内折射率和细胞厚度均相关。当用阿霉素和氯喹这两种产生不同表型的细胞毒性化合物处理时，DHM能够区分两种代表性细胞系（HeLa和H9c2）的细胞形态变化。我们在终点测量中分析了与细胞形态和细胞内含量相关的参数，并通过延时记录进一步研究了这些参数。将DHM获得的结果与其他无标记光学显微镜技术进行了比较，即相衬显微镜、微分干涉相衬显微镜和强度传输方程（由三张明场图像重建）。为了进行比较，在不同的电动显微镜上以常见的96孔板格式采集图像。与所检测的其他显微镜不同，使用DHM生成的图像可以通过一种简单的自动实时分析方法轻松定量，以区分每个细胞系中产生的不同表型。DHM技术适用于开发强大且无偏差的基于图像的检测方法。
