The Laser Scanning Confocal Microscope Cell Image Processing And The Cell 3D Parameters Stereological Quantitative Analysis

Stereology is an interdisciplinary method for 3D morphological study developed from mathematics and morphology. Stereology, which is a fast, reliable, unbiased and high repetitive analysis method, analyzes the objects 3D information through the 2D image measurement. This paper proposes a novel quantitative analysis method for cell that is combining the stereology and laser scanning confocal microscope (LSCM) imaging.Before the measurement, we give an introduction of LSCM images pre-processing which includes the noise filter, pseudo-color pimple separation, fault images rendering. Besides, we also study about the feasibility of stereological measurement on LSCM images. Image information extraction method, edge correction method and point matching interpolation method are compared with stereology by measuring 3D parameters of the same set of LSCM cell images, the results show that the stereology method is an acceptable way for cell images quantitative analysis.After approving the feasibility of stereological measurement, three groups of B16F10, B16/Vector and B16/GPR4 cells suspended in culture media are studied and 20 image sets of each are taken and used in the experiments. The results show that the control group B16/Vector has no significant differences as compared with the parental B16F10 cells, while the experimental group B16/GPR4 shows significant differences in the stereological parameters of the cellular surface area to volume ratio (Rsvc) and the relative cellular form factor (FFc) (p<0.05 for Rsvc, and p<0.02 for FFc, respectively), suggesting that B16/GPR4 cells have less surface protrusions and reduced irregularity. This is in line with the previous observations that GPR4 inhibits tumor cell migration and invasion in the transwell assay.At last, the system which is based on MATLAB is introduced amply. The system is composed by image basic edition, images pro-processing, images interpolation and edge correction, stereological 3D parameter measure. It provides a reliable and convenient tool for cell quantitative analysis.