Rappaz Benjamin, Marquet Pierre, Cuche Etienne, Emery Yves, Depeursinge Christian, Magistretti Pierre
Opt Express. 2005 Nov 14;13(23):9361-73. doi: 10.1364/opex.13.009361.
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 to lambda/1800, and the low acquisition time (~20micros) 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.
我们开发了一种透射模式的数字全息显微镜（DHM），适用于细胞动力学的定量研究。通过测量培养中的活细胞在透射波前产生的相移来对其进行光学探测。相位信号的高时间稳定性（相当于λ/1800）和较短的采集时间（约20微秒）使得能够监测细胞动力学过程。本文提出了一种从测量的相移计算积分折射率和细胞厚度（形态测量）的实验方法。具体而言，该方法已应用于研究低渗应激期间培养神经元的动力学。这种应激会导致相位出现反常下降，而通过应用本文所述的方法可以完全解决这一问题；实际上，该方法能够独立确定细胞的厚度和积分折射率。
