Automatic image-based method for quantitative analysis of photosynthetic cell cultures

Abstract

This work deals with an automatic quantitative analysis of photosynthetic cell cultures. It uses images captured by a confocal fluorescent microscope for automatic determination the number of cells in sample containing complex 3D structure of cell clusters. Experiments were performed on the confocal microscope Leica TCS SP8 X. The cell nuclei were stained by SYBR® Green fluorescent DNA binding marker. In the first step we used combination of adaptive thresholding to found out areas where nuclei were located. Proposed segmentation steps allowed reduction of noise and artefacts. Z-axis position was obtained as a location of peak from intensity profile. Finally model of scene can be created by emplacement of spheres with adequate diameter to found 3D coordinates. Number of cells per volumetric unit were determined in structurally different culture samples of cell cultures Chenopodium rubrum (Cr) and Solanum lycopersicum (To). The results were verified by manual counting.