New Phytologist 2010-06-01

Perfluorodecalin enhances in vivo confocal microscopy resolution of Arabidopsis thaliana mesophyll.

George R Littlejohn, João D Gouveia, Christoph Edner, Nicholas Smirnoff, John Love

Index: New Phytol. 186(4) , 1018-25, (2010)

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Abstract

*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.