Ecole Polytechnique Fédérale de Lausanne, Laboratory of Neuroenergetics and Cellular Dynamics, Lausanne, Switzerland.
J Biomed Opt. 2013 Mar;18(3):036007. doi: 10.1117/1.JBO.18.3.036007.
A dual-wavelength digital holographic microscope to measure absolute volume of living cells is proposed. The optical setup allows us to reconstruct two quantitative phase contrast images at two different wavelengths from a single hologram acquisition. When adding the absorbing dye fast green FCF as a dispersive agent to the extracellular medium, cellular thickness can be univocally determined in the full field of view. In addition to the absolute cell volume, the method can be applied to derive important biophysical parameters of living cells including osmotic membrane water permeability coefficient and the integral intracellular refractive index (RI). Further, the RI of transmembrane flux can be determined giving an indication about the nature of transported solutes. The proposed method is applied to cultured human embryonic kidney cells, Chinese hamster ovary cells, human red blood cells, mouse cortical astrocytes, and neurons.
提出了一种用于测量活细胞绝对体积的双波长数字全息显微镜。该光学设置允许我们从单个全息图采集重建两个不同波长的定量相衬图像。当将吸收染料 Fast Green FCF 添加到细胞外介质中作为分散剂时，可以在整个视场中唯一确定细胞厚度。除了绝对细胞体积外，该方法还可用于推导活细胞的重要生物物理参数，包括渗透膜水渗透率系数和细胞内整体折射率（RI）。此外，还可以确定跨膜通量的 RI，从而提供有关转运溶质性质的信息。该方法应用于培养的人胚肾细胞、中国仓鼠卵巢细胞、人红细胞、小鼠皮质星形胶质细胞和神经元。
