Fig. 5
Principal steps of mast cell quantification protocol using ImageJ software program. a Original photo illustrating the subepidermal layer of a one-day old skin wound on the limb stained with the CD117 antibody. The blue line represents the border between the subepidermal and the deeper dermal layer. b Staining of CD117+ mast cells after colour deconvolution and c its binary transformation. d Manual subtraction of stained cutaneous appendages; the deeper dermal region below the blue line was also deleted. e Remaining cells stained by the antibody; blue cells are those included in the count by the software (cell area between 8 and 300 μm2). The original photo was taken at a 200X magnification. Arrow head points to a stained mast cell