title: Perfluorodecalin enhances in vivo confocal microscopy resolution of Arabidopsis thaliana mesophyll.
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.
authors.affiliation: School of Biosciences, The University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK.
authors.forename: George R
authors.initials: GR
authors.lastname: Littlejohn
authors.affiliation: School of Biosciences, The University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK.
authors.forename: João D
authors.initials: JD
authors.lastname: Gouveia
authors.affiliation: School of Biosciences, The University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK.
authors.forename: Christoph
authors.initials: C
authors.lastname: Edner
authors.affiliation: School of Biosciences, The University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Smirnoff
authors.affiliation: School of Biosciences, The University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK.
authors.forename: John
authors.initials: J
authors.lastname: Love
mesh_terms: D017360:Arabidopsis / Q000166:cytology* / Q000187:drug effects; D001692:Biological Transport / Q000187:drug effects; D005466:Fluorocarbons / Q000494:pharmacology*; D018525:Germination / Q000187:drug effects; D021621:Imaging, Three-Dimensional; D018613:Microscopy, Confocal / Q000379:methods*; D045332:Photosystem II Protein Complex / Q000378:metabolism; D018515:Plant Leaves / Q000166:cytology* / Q000187:drug effects* / Q000502:physiology; D036226:Seedlings / Q000187:drug effects / Q000254:growth & development; D013050:Spectrometry, Fluorescence
chemical_list: D005466:Fluorocarbons; D045332:Photosystem II Protein Complex; C008805:perfluorodecalin