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Dynamic quantitative analysis of adherent cell cultures by means of lens-free video microscopy

机译:无透镜视频显微镜通过透镜视频显微镜动态定量分析粘附细胞培养物

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We present our implementation of lens-free video microscopy setup for the monitoring of adherent cell cultures. We use a multi-wavelength LED illumination together with a dedicated holographic reconstruction algorithm that allows for an efficient removal of twin images from the reconstructed phase image for densities up to those of confluent cell cultures(>500 cells/mm~2). We thereby demonstrate that lens-free video microscopy, with a large field of view(~30 mm~2)can enable us to capture the images of thousands of cells simultaneously and directly inside the incubator. It is then possible to trace and quantify single cells along several cell cycles. We thus prove that lens-free microscopy is a quantitative phase imaging technique enabling estimation of several metrics at the single cell level as a function of time, for example the area, dry mass, maximum thickness, major axis length and aspect ratio of each cell. Combined with cell tracking, it is then possible to extract important parameters such as the initial cell dry mass(just after cell division), the final cell dry mass(just before cell division), the average cell growth rate, and the cell cycle duration. As an example, we discuss the monitoring of a HeLa cell cultures which provided us with a data-set featuring more than 10 000 cell cycle tracks and more than 2x10~6 cell morphological measurements in a single time-lapse.
机译:我们展示了我们对粘附细胞培养物监测的无透镜视频显微镜设置的实现。我们使用多波长LED照明以及专用全息重建算法,其允许从重建的相位图像中有效地去除双图像,以便密度高达汇合细胞培养物(> 500个细胞/ mm〜2)。因此,我们证明了无透镜的视频显微镜,具有大视野(〜30mm〜2),可以使我们同时捕获成千上万个体的图像,并直接在孵化器内。然后可以沿几种细胞循环追踪和量化单个细胞。因此,我们证明了无透镜显微镜是定量相位成像技术,可实现单个电池水平的几个度量,例如时间,例如区域,干料,最大厚度,主轴长度和每个单元的纵横比。结合细胞跟踪,然后可以提取重要的参数,例如初始细胞干肿块(刚性细胞分裂后),最终细胞干含量(在细胞分裂之前),平均细胞生长速率和细胞周期持续时间。作为一个例子,我们讨论了监测HeLa细胞培养物,其提供了具有超过10 000个细胞周期轨迹的数据集,并且在单一的时间流逝中具有超过2×10〜6个细胞形态测量。

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