Kastl Lena, Isbach Michael, Dirksen Dieter, Schnekenburger Jürgen, Kemper Björn
Biomedical Technology Center, University of Muenster, Mendelstr. 17, Muenster, D-48149, Germany.
Department of Prosthetic Dentistry and Biomaterials, University of Muenster, Waldeyerstraße 30, Muenster, D-48149, Germany.
Cytometry A. 2017 May;91(5):470-481. doi: 10.1002/cyto.a.23082. Epub 2017 Mar 6.
The potential of quantitative phase imaging (QPI) with digital holographic microscopy (DHM) for quantification of cell culture quality was explored. Label-free QPI of detached single cells in suspension was performed by Michelson interferometer-based self-interference DHM. Two pancreatic tumor cell lines were chosen as cellular model and analyzed for refractive index, volume, and dry mass under varying culture conditions. Firstly, adequate cell numbers for reliable statistics were identified. Then, to characterize the performance and reproducibility of the method, we compared results from independently repeated measurements and quantified the cellular response to osmolality changes of the cell culture medium. Finally, it was demonstrated that the evaluation of QPI images allows the extraction of absolute cell parameters which are related to cell layer confluence states. In summary, the results show that QPI enables label-free imaging cytometry, which provides novel complementary integral biophysical data sets for sophisticated quantification of cell culture quality with minimized sample preparation. © 2017 International Society for Advancement of Cytometry.
探索了利用数字全息显微镜（DHM）进行定量相成像（QPI）以量化细胞培养质量的潜力。基于迈克尔逊干涉仪的自干涉DHM对悬浮液中分离的单个细胞进行无标记QPI。选择两种胰腺肿瘤细胞系作为细胞模型，在不同培养条件下分析其折射率、体积和干质量。首先，确定了进行可靠统计所需的足够细胞数量。然后，为了表征该方法的性能和可重复性，我们比较了独立重复测量的结果，并量化了细胞对细胞培养基渗透压变化的反应。最后，证明对QPI图像的评估能够提取与细胞层汇合状态相关的绝对细胞参数。总之，结果表明QPI能够实现无标记成像细胞术，为复杂的细胞培养质量量化提供了新的互补性完整生物物理数据集，且样品制备最少。© 2017国际细胞计量学会。
