Perfluorodecalin enhances in vivo confocal microscopy resolution of Arabidopsis thaliana mesophyll.
*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.
Littlejohn
George R
GR
School of Biosciences, The University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK.
Gouveia
João D
JD
School of Biosciences, The University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK.
Edner
Christoph
C
School of Biosciences, The University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK.
Smirnoff
Nicholas
N
School of Biosciences, The University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK.
Love
John
J
School of Biosciences, The University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK.
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
D005466:Fluorocarbons; D045332:Photosystem II Protein Complex; C008805:perfluorodecalin