Sequential processing of quantitative phase images for the study of cell behaviour in real‐time digital holographic microscopy
Sequential processing of quantitative phase images for the study of cell behaviour in real‐time digital holographic microscopy
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DOI：
10.1111/jmi.12165
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发表时间：
2014-11
影响因子：
2
通讯作者：
Tomáš Zikmund;Lukáš Kvasnica;Matěj Týč;A. Křížová;J. Čolláková;R. Chmelík
中科院分区：
文献类型：
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作者：
Tomáš Zikmund;Lukáš Kvasnica;Matěj Týč;A. Křížová;J. Čolláková;R. Chmelík
透射光全息显微镜特别用于活细胞等透明微观物体的定量相位成像。细胞的研究基于从相位图像的延时序列中提取细胞行为的动态数据。然而，相位图像受到相位像差的影响，这使得分析变得特别困难。这是因为在长期实验过程中相变形容易发生变化。在这里，我们提出了一种新颖的算法，用于以延时序列顺序处理活细胞相位图像。该算法使用加权最小二乘曲面拟合来补偿相位图像的变形。此外，它还可以识别并分割相位图像中的单个细胞。所有这些程序都是自动执行的，并在获得每个单相图像后立即应用。该算法的这一特性对于实时细胞定量相成像以及通过回放记录序列直至实际时间来瞬时控制实验过程非常重要。这种操作员的干预是源自图像分析的过程自动化的先驱。使用离轴全息显微镜在大鼠纤维肉瘤细胞图像上证明了所提出算法的效率。
Transmitted light holographic microscopy is particularly used for quantitative phase imaging of transparent microscopic objects such as living cells. The study of the cell is based on extraction of the dynamic data on cell behaviour from the time‐lapse sequence of the phase images. However, the phase images are affected by the phase aberrations that make the analysis particularly difficult. This is because the phase deformation is prone to change during long‐term experiments. Here, we present a novel algorithm for sequential processing of living cells phase images in a time‐lapse sequence. The algorithm compensates for the deformation of a phase image using weighted least‐squares surface fitting. Moreover, it identifies and segments the individual cells in the phase image. All these procedures are performed automatically and applied immediately after obtaining every single phase image. This property of the algorithm is important for real‐time cell quantitative phase imaging and instantaneous control of the course of the experiment by playback of the recorded sequence up to actual time. Such operator's intervention is a forerunner of process automation derived from image analysis. The efficiency of the propounded algorithm is demonstrated on images of rat fibrosarcoma cells using an off‐axis holographic microscope.