The monitoring of the dynamics of stem cells’ growth in culture is important in regenerative medicine. In this paper the method of cells’ images segmentation based on alternating microscopic imaging with bright field (BF) and epifluorescent (EF) images is proposed. The method consists of two principal stages: coarse segmentation of EF images followed by fine segmentation on BF ones. The latter is based on the morphological watershed from markers produced in the first stage. Due to the fact that sequence of EF is shorter than BF one, markers cannot be produced directly for all BF images. In order to create them, an additional step of morphological interpolation of markers is applied.