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Perfluorodecalin enhances in vivo confocal microscopy resolution of Arabidopsis thaliana mesophyll
Article first published online: 29 MAR 2010
DOI: 10.1111/j.1469-8137.2010.03244.x
© The Authors (2010). Journal compilation © New Phytologist Trust (2010)
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How to Cite
Littlejohn, G. R., Gouveia, J. D., Edner, C., Smirnoff, N. and Love, J. (2010), Perfluorodecalin enhances in vivo confocal microscopy resolution of Arabidopsis thaliana mesophyll. New Phytologist, 186: 1018–1025. doi: 10.1111/j.1469-8137.2010.03244.x
Publication History
- Issue published online: 10 MAY 2010
- Article first published online: 29 MAR 2010
- Received: 17 December 2009, Accepted: 14 February 2010
Keywords:
- Arabidopsis;
- confocal microscopy;
- fluorescence;
- mesophyll;
- perfluorocarbon;
- perfluorodecalin
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.