A simple morphometric analysis method for dermal microstructure using color thresholding and moments

Abstract Background Proper assessment of dermal collagen fibers by dermatologists and researchers is essential. Histologic evaluation methods have limitations. We present a simple method for measurement of collagen fibers in human skin using Masson's trichrome staining. Materials and Methods Normal skin specimens from a cadaver were processed with Masson's trichrome, which can effectively stain collagen fibers blue with aniline dye. Optical photomicrographs of these slides were analyzed using ImageJ software. Color image processing, a histogram‐based function of ImageJ for image segmentation, was performed with color moments thresholding technique. We selected blue areas by adjusting the blue channel to include specific values. The selected areas were highlighted and evaluated. We divided the image into layers of 0.09‐mm 2 areas from the top to bottom of the dermis. Each area was cropped and evaluated. Results Quantitative assessment yielded the quantitative size occupied by collagen fibers in an area of 0.09 mm 2 . Calculation of the percentage in each area can be used to determine the density of collagen fibers. Conclusion Measurements obtained with our method can be applied to research on dermal collagen fibers. We present a convenient quantitative assessment method for the dermal constituents in Masson's trichrome‐stained slides.