Observer z1 spinning disc confocal microscope

Trichrome and immunohistochemistry stained sections were observed using a Zeiss Axiovert 200 light microscope at 40X magnification, e.g., high power field (HPF). Immunofluorescence images were taken through a Zeiss observer Z1 spinning disc confocal microscope. The size of granulation tissues was quantified according to trichrome stained whole wound sections. Quantification of collagen deposition was performed on trichrome stained HPF images. Quantification of infiltrated cell number was performed on five DAPI stained HPF images from the center of the scaffold areas in the wound sections for each group. Quantification of angiogenesis was performed by counting the number and the size of vessels per HPF. Proportions of F4/80 positive cells either from the scaffolds or from the wound beds were calculated respectively based on five representatives images in each group. Proportions of ASM positive cells from the center of the scaffold areas of the histological sections or from in vitro HDF seeded scaffolds were determined from nine representative images each group. Quantification was performed blindly and with the assistant of ImageJ software. Data are presented as mean ± SEM.