Measurement of the integral refractive index and dynamic cell morphometry of living cells with digital holographic microscopy
Measurement of the integral refractive index and dynamic cell morphometry of living cells with digital holographic microscopy
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DOI：
10.1364/opex.13.009361
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发表时间：
2005-11-14
期刊：
影响因子：
3.8
通讯作者：
Magistretti, PJ
中科院分区：
文献类型：
--
作者：
Rappaz, B;Marquet, P;Magistretti, PJ
我们已经开发了一个数字全息显微镜（DHM），在传输模式，适合于细胞动力学的定量研究。培养中的活细胞通过测量它们在透射波前上产生的相移来进行光学探测。相位信号的高时间稳定性（等效λ/1800）和低采集时间（类似于20 μ s）使得能够监测细胞动力学过程。一个实验程序，允许计算的积分折射率和细胞厚度（形态学）从测得的相移。具体而言，该方法已被应用于研究在培养过程中的低渗应力的神经元的动力学。这种压力产生一个矛盾的减少的阶段，可以完全解决通过应用本文中所描述的方法的方法;事实上，该方法允许独立地确定的厚度和积分折射率的细胞。(c)2005年，美国光学学会。
We have developed a digital holographic microscope (DHM), in a transmission mode, adapted to the quantitative study of cellular dynamics. Living cells in culture are optically probed by measuring the phase shift they produce on the transmitted wave front. The high temporal stability of the phase signal, equivalent lambda/1800, and the low acquisition time (similar to 20 mu s) enable to monitor cellular dynamics processes. An experimental procedure allowing to calculate both the integral refractive index and the cellular thickness (morphometry) from the measured phase shift is presented. Specifically, the method has been applied to study the dynamics of neurons in culture during a hypotonic stress. Such stress produces a paradoxical decrease of the phase which can be entirely resolved by applying the methodological approach described in this article; indeed the method allows to determine independently the thickness and the integral refractive index of cells. (c) 2005 Optical Society of America.