Quantitative stereological analysis of confocal laser scanning microscopic images of cells

Stereology is an interdisciplinary method of mathematics and morphology, and is unbiased, quick, accurate, noninvasive, and has high reproducibility. It is widely used in radiation imaging and cell biology. In this paper, the confocal laser scanning microscopic images (CLSMI) of tumor cells of MCF-7 and B16/ GPR4 were quantitatively analyzed using stereological point counting method. 22 images of MCF-7 and 16 images of B16/GPR4 were used in the study. The number of slices used to space the cell was determined by keeping the coefficient of error (CE) equal or less than 5%. The cell was sliced by randomly picking up the starting point and the stereology grids were placed in the 2D images of the cell layers. The circumference, areas and volumes of the cells and nuclei were measured. The average volume of nuclei (Vn), volume density of nuclei (Vvn), surface area density of nuclei (Svn), ratio of surface area and volume of nuclei (Rsvn), and ratio of nuclei and plasma (Rnp) were calculated. The maximum and minimum, and median diameters of the cells were calculated from the binarized image of the cells. The results were very close to the results obtained using the image processing software of Leica and reconstructed 3D images of the CLSMI. It was shown that the stereological method is a quick and easy-to-use tool for quantitative analysis of cell images.