Refractive index measurement in viable cells using quantitative phase-amplitude microscopy and confocal microscopy
Refractive index measurement in viable cells using quantitative phase-amplitude microscopy and confocal microscopy
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DOI：
10.1002/cyto.a.20134
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发表时间：
2005-05-01
期刊：
影响因子：
3.7
通讯作者：
Delbridge, LMD
中科院分区：
文献类型：
--
作者：
Curl, CL;Bellair, CJ;Delbridge, LMD
背景：细胞材料的折射率（RI）提供有关细胞组成和组织结构的基本生物物理信息。描述细胞折射特性的努力因测量活细胞 RI 时遇到的实验困难而受到严重阻碍。在本报告中，我们描述了应用定量相位显微镜与共焦显微镜结合测量培养的肌肉细胞样本的 RI 的程序。方法：实验策略包括使用共焦光学切片程序计算细胞厚度，使用定量相位显微镜构建同一细胞的相位图，以及选择感兴趣的细胞区域来求解细胞 RI。结果：平均细胞确定了六个细胞区域（五个细胞质和一个细胞核）的厚度和相位值。计算出的细胞质和核区域的平均折射率为1.360±0.004。最终 RI 值的不确定性代表了技术测量误差。结论：我们描述的使用该原型细胞进行活细胞 RI 测量的方法在细胞生长和功能反应的研究中具有广泛的通用应用。我们报告的 RI 值可用于培养细胞结构和形态的光学分析。 (c) 2005 年 Wiley-Liss, Inc.
Background: The refractive index (RI) of cellular material provides fundamental biophysical information about the composition and organizational structure of cells. Efforts to describe the refractive properties of cells have been significantly impeded by the experimental difficulties encountered in measuring viable cell RI In this report we describe a procedure for the application of quantitative phase microscopy in conjunction with confocal microscopy to measure the RI of a cultured muscle cell specimen.Methods: The experimental strategy involved calculation of cell thickness by using confocal optical sectioning procedures, construction of a phase map of the same cell using quantitative phase microscopy, and selection of cellular regions of interest to solve for the cell RI.Results: Mean cell thickness and phase values for six cell regions (five cytoplasmic and one nuclear) were determined. The average refractive index calculated for cytoplasmic and nuclear regions was 1.360 0.004. The uncertainty in the final RI value represents the technique measurement error.Conclusions: The methodology we describe for viable cell RI measurement with this prototype cell has broad generic application in the study of cell growth and functional responses. The RI value we report may be used in optical analyses of cultured cell structure and morphology. (c) 2005 Wiley-Liss, Inc.