Digital Image Analysis: Cell & Nuclei Count

 

Cultured endothelial cells (Monolayer) were stained with calcein-AM (cytosol, green color) and Hoechst (nuclei, blue color) and digital images were analysed by counting either cells or nuclei after separating the color channels.
monolayer
cell count (green channel) nuclei count (blue channel)
monolyaer cells
monolayer nuclei
cells after segmentation nuclei after segmentation
watershed cells
watershed nuclei
53 cells counted 54 nuclei counted
outlines cells
outlines nuclei

The colored image was splitted in RGB channels, the green and blue channels were thresholded and objects were counted after morphological modification. Objects touching the edge of the image were excluded, 53 cells and 54 nuclei were counted. The difference accounts for a cell touching the edge in the upper left while the nucleus does not.