Loparev A V, Kretushev A V, Tychinskiĭ V P
Biofizika. 2008 Mar-Apr;53(2):299-304.
A new method for the identification of intracellular structures of a living cell and obtaining the quantitative parameters characterizing these structures by means of coherent phase microscopy is proposed. The method is based on the analysis of the histogram of a cell phase image and its decomposition by phase height levels. In the spherical and cylindrical approximation of the cell, the method makes it possible to separate the contributions of phase-contrast intracellular structures to the integral refractive index of the whole cell. The calculation of refractive indices of intracellular structures is illustrated on a two-component model of a spherical cell. The possibility of determining the refractive indices of cellular organelles was shown by an example of cyanobacterium Anabaena variabilis ATCC 29413 cells and Cancer mammae breast cancer cells. Three most contrast cellular structures in the phase images of the cells were identified, and their refractive indices were determined. For Anabaena variabilis cells, these structures were the cell wall (<n(cw)> = 1.39), tylakoids (<n(T)> = 1.47), and the nucleoid (<n(nucl)> = 1.58); for breast cancer cells, these were the cytoplasm (<n(cyt)> = 1.34), the nucleus (<n(n)> = 1.36), and the nucleolus (<n(nucleol)> = 1.44). Using this method is possible to identify phase-contrast intracellular structures and calculate their refractive indices. This enables one to follow morphofunctional changes of not only the whole cell but also its individual organelles.
提出了一种通过相干相显微镜识别活细胞内结构并获取表征这些结构的定量参数的新方法。该方法基于对细胞相位图像直方图的分析及其按相位高度水平的分解。在细胞的球形和柱形近似中，该方法能够分离相差细胞内结构对整个细胞积分折射率的贡献。在球形细胞的双组分模型上说明了细胞内结构折射率的计算。以多变鱼腥藻ATCC 29413细胞和乳腺癌细胞为例，展示了确定细胞器折射率的可能性。确定了细胞相位图像中三个对比度最高的细胞结构，并测定了它们的折射率。对于多变鱼腥藻细胞，这些结构是细胞壁（<n(cw)> = 1.39）、类囊体（<n(T)> = 1.47）和拟核（<n(nucl)> = 1.58）；对于乳腺癌细胞，这些是细胞质（<n(cyt)> = 1.34）、细胞核（<n(n)> = 1.36）和核仁（<n(nucleol)> = 1.44）。使用该方法可以识别相差细胞内结构并计算它们的折射率。这使得人们不仅能够追踪整个细胞的形态功能变化，还能追踪其单个细胞器的形态功能变化。
