Quantification of Melanosome Transfer Using Immunofluorescence Microscopy and Automated Image Analysis

Methods Mol Biol. 2020:2109:55-65. doi: 10.1007/7651_2019_241.

Abstract

The study of skin pigmentation requires determining the rate of melanin production in melanocytes and quantifying the rate of melanosome transfer to keratinocytes. Here, we describe a method to quantify melanosome transfer using immunofluorescence microscopy coupled with automated image analysis of in vitro human melanocytes and keratinocytes in co-culture. In this method, the number of melanin capped keratinocyte nuclei is quantified.

Keywords: Image quantification; Melanin transfer quantitation; Melanocyte keratinocyte co-culture; Melanosome transfer.

MeSH terms

  • Cells, Cultured
  • Coculture Techniques
  • Humans
  • Keratinocytes / cytology*
  • Keratinocytes / metabolism
  • Melanins / metabolism
  • Melanocytes / cytology*
  • Melanocytes / metabolism
  • Melanosomes / metabolism
  • Melanosomes / transplantation*
  • Microscopy, Fluorescence
  • Radiographic Image Interpretation, Computer-Assisted

Substances

  • Melanins