Quantitative phase microscopy: a new tool for measurement of cell culture growth and confluency in situ
Quantitative phase microscopy: a new tool for measurement of cell culture growth and confluency in situ
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DOI：
10.1007/s00424-004-1248-7
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发表时间：
2004-07-01
影响因子：
4.5
通讯作者：
Delbridge, LMD
中科院分区：
文献类型：
--
作者：
Curl, CL;Harris, T;Delbridge, LMD
定量相位显微镜（QPM）是最近开发的一种计算方法，提供定量相位测量的样品图像下获得的明场条件下，没有相位或干涉衬度光学。为了执行QPM，获取焦点对准的明场图像以及一个正散焦图像和一个负散焦图像。然后应用算法来产生样本相位图。在这项调查中，我们表明，操纵的相位图强度直方图，使用新的，非主观的阈值和分割方法提供了增强的描绘细胞培养。QPM用于测量培养的气道平滑肌细胞在92小时内的生长行为。在这段时间内，平行的QPM导出的汇合测量值和血细胞计数导出的气道平滑肌细胞计数之间存在高度相关性。使用QPM，半透明细胞可以可视化，具有改进的细胞边界定义，允许精确和可重复地测量细胞培养物汇合。定量相位成像提供了一种快速、光学简单和非破坏性的方法来测量细胞形态。基于QPM的分析方法的进一步发展有可能提供更精确的细胞生长测量。
Quantitative phase microscopy (QPM) is a recently developed computational approach that provides quantitative phase measurements of specimen images obtained under bright-field conditions without phase- or interference-contrast optics. To perform QPM, an in-focus bright-field image is acquired, together with one positive and one negative de-focus image. An algorithm is then applied to produce a specimen phase map. In this investigation we demonstrate that manipulation of the phase map intensity histogram using novel, non-subjective thresholding and segmentation methods provides enhanced delineation of cells in culture. QPM was utilised to measure the growth behaviour of cultured airway smooth muscle cells over a 92-h period. There was a high degree of correlation between parallel QPM-derived confluency measurements and haemocytometry-derived counts of airway smooth muscle cells over this time period. Using QPM, translucent cells can be visualised with improved cell boundary definition allowing precise and reproducible measurements of cell culture confluency. Quantitative phase imaging provides a rapid, optically simple and non-destructive approach for measurement of cellular morphology. Further development of the QPM-based analysis methodology has the potential to provide even more refined measures of cellular growth.