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

Ji Hyuck Hong, Dai Hyun Kim, Im Joo Rhyu, Young Chul Kye, Hyo Hyun Ahn

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)


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-mm2 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 mm2. 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.

Original languageEnglish
Pages (from-to)132-136
Number of pages5
JournalSkin Research and Technology
Issue number1
Publication statusPublished - 2020 Jan 1

Bibliographical note

Funding Information:
This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Science, ICT & Future Planning (NRF‐2017R1A2B4011001).

Publisher Copyright:
© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd


  • Histology
  • ImageJ
  • Masson's trichrome
  • cadaver
  • collagen fiber
  • morphometric analysis

ASJC Scopus subject areas

  • Dermatology


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