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Perfluorodecalin enhances in vivo confocal microscopy resolution of Arabidopsis thaliana mesophyll
Author(s) -
Littlejohn George R.,
Gouveia João D.,
Edner Christoph,
Smirnoff Nicholas,
Love John
Publication year - 2010
Publication title -
new phytologist
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.742
H-Index - 244
eISSN - 1469-8137
pISSN - 0028-646X
DOI - 10.1111/j.1469-8137.2010.03244.x
Subject(s) - confocal , confocal microscopy , vascular bundle , biophysics , arabidopsis thaliana , confocal laser scanning microscopy , microscopy , resolution (logic) , biology , fluorescence , chemistry , botany , microbiology and biotechnology , optics , biochemistry , physics , artificial intelligence , gene , mutant , computer science
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.