For successful tissue engineering, an even and stable layer of the transplanted cells must be made on a porous scaffold. However, uneven cell seeding and unstable positioning of the transplanted cells on the scaffold can cause the loss of transplanted cells and the intrusion of other kinds of cells, precluding the formation of a uniform and stable layer of transplanted cells. Therefore cell sheet have to be developed and also distribution of the seeded cells on sheet should be identified for making a uniform and stable cell layer. Several methods can be used to make cell sheets and confirm the cell distributions, including the use of microscopy and paraffin sections, but they are either difficult to use or display only very thin and small parts of the scaffold. In this study, we made cell sheets from poly(D,L-lactide-co-glycolide) solution and dermis tissue, and identified the distribution status of all the cells using a bioimage analyzer. We obtained a result that the two sheets were good for making uniform cell layer and the Bioimage analyzer was very useful for identifying overall distribution status of the cells.