Development of intravital intermittent confocal imaging system for studying Langerhans cell turnover

J Invest Dermatol. 2006 Nov;126(11):2452-7. doi: 10.1038/sj.jid.5700448. Epub 2006 Jun 22.


Although several studies have suggested relatively slow turnover of Langerhans cells (LCs), their actual lifespan remains elusive. Here we report the development of a new intravital imaging system for studying LC efflux and influx. Epidermal LCs expressing enhanced green fluorescent protein (EGFP) were visualized in anesthetized I-Abeta-EGFP knock-in mice by confocal microscopy. By overlaying two sets of EGFP+ LC images recorded in the same microscopic fields at time 0 and 24 hours later, we identified LC subpopulations that had disappeared from or newly emerged in the epidermis during that period. Of >10,000 LCs analyzed in this manner, an overwhelming majority (97.8+/-0.2%) of LCs showed no significant changes in the x-y locations, whereas 1.3+/-0.1% of the LCs that were found at time 0 became undetectable 24 hours later, representing LC efflux. Conversely, 0.9+/-0.1% of the LCs that were found at time 24 hours were not detectable at time 0, representing LC influx. From these frequencies, we estimated the half-life of epidermal LCs to range from 53 to 78 days, providing new insights into the immunobiology of LCs. Our intermittent imaging approach may be regarded as a technical breakthrough enabling direct visual assessment of LC turnover in living animals.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Epidermal Cells*
  • Green Fluorescent Proteins / analysis*
  • Green Fluorescent Proteins / genetics
  • Langerhans Cells / chemistry
  • Langerhans Cells / cytology*
  • Mice
  • Mice, Mutant Strains
  • Microscopy, Confocal / methods*


  • enhanced green fluorescent protein
  • Green Fluorescent Proteins