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Keywords:

  • Arabidopsis;
  • confocal microscopy;
  • fluorescence;
  • mesophyll;
  • perfluorocarbon;
  • perfluorodecalin

Summary

  • Air spaces in the leaf mesophyll generate deleterious optical effects that compromise confocal microscopy.
  • Leaves were mounted in the nontoxic, nonfluorescent perfluorocarbon, perfluorodecalin (PFD), and optical enhancement and physiological effect were assessed using confocal microscopy and chlorophyll fluorescence.
  • Mounting leaves of Arabidopsis thaliana in PFD significantly improved the optical qualities of the leaf, thereby enabling high-resolution laser scanning confocal imaging over twofold deeper into the mesophyll, compared with using water. Incubation in PFD had less physiological impact on the mounted specimen than water.
  • We conclude that the application of PFD as a mounting medium substantially increases confocal image resolution of living mesophyll and vascular bundle cells, with minimal physiological impact.