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Quantification of Melanosome Transfer Using Immunofluorescence Microscopy and Automated Image Analysis

  • Aishwarya Sridharan
  • S. Y. John Lim
  • Graham D. Wright
  • Leah A. VardyEmail author
Protocol
  • 927 Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 2109)

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

Melanin transfer quantitation Melanosome transfer Melanocyte keratinocyte co-culture Image quantification 

Supplementary material

472479_4_En_241_MOESM1_ESM.zip (6 kb)
Pipeline.cpproj

References

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    Meredith P, Sarna T (2006) The physical and chemical properties of eumelanin. Pigment Cell Res 19(6):572–594CrossRefGoogle Scholar
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    Dickson MA et al (2000) Human keratinocytes that express hTERT and also bypass a p16(INK4a)-enforced mechanism that limits life span become immortal yet retain normal growth and differentiation characteristics. Mol Cell Biol 20(4):1436–1447CrossRefGoogle Scholar
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    Kamentsky L et al (2011) Improved structure, function and compatibility for cell profiler: modular high-throughput image analysis software. Bioinformatics 27(8):1179–1180CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2019

Authors and Affiliations

  • Aishwarya Sridharan
    • 1
  • S. Y. John Lim
    • 1
  • Graham D. Wright
    • 1
  • Leah A. Vardy
    • 1
    • 2
    Email author
  1. 1.Skin Research Institute of Singapore, A*STARSingaporeSingapore
  2. 2.School of Biological SciencesNanyang Technological UniversitySingaporeSingapore

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