Perfluorodecalin enhances in vivo confocal microscopy resolution of Arabidopsis thaliana mesophyll
2010/03/29 by George R. Littlejohn, João D. Gouveia, Joao Gouveia +3 · 96 citations
Agricultural and Biological Sciences · Biochemistry, Genetics and Molecular Biology · Chemistry · #Advanced Fluorescence Microscopy Techniques #Arabidopsis thaliana #Biochemistry #Biocrusts and Microbial Ecology #Biology #Biophysics #Botany #Cell biology #Chemistry #Confocal #Confocal laser scanning microscopy #Confocal microscopy #Fluorescence #Microscopy #Optics #Photosynthetic Processes and Mechanisms #Resolution (logic) #Vascular bundle
paper · pdf · doi:10.1111/j.1469-8137.2010.03244.x
published in New Phytologist 186(4), 1018-1025 (Wiley)
openalex publication_date 2010/03/29 · openalex created_date 2016/06/24 · openalex updated_date 2026/07/28
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.
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