2010/03/29 by George R. Littlejohn, João D. Gouveia, Joao Gouveia +3 · 4 citations
Biochemistry, Genetics and Molecular Biology · Agricultural and Biological Sciences · #Photosynthetic Processes and Mechanisms #Advanced Fluorescence Microscopy Techniques #Biocrusts and Microbial Ecology
paper · pdf · doi:10.1111/j.1469-8137.2010.03244.x
*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.