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Botany JoVE (Open Access) Citable · DOI

A Simple Method for Imaging Arabidopsis Leaves Using Perfluorodecalin as an Infiltrative Imaging Medium

DOI: 10.3791/3394-v
What you'll learn
  • ✓ Mount Arabidopsis leaves in perfluorodecalin for improved optical imaging depth
  • ✓ Acquire high-resolution confocal/fluorescence images of leaf tissue with minimal phototoxicity
  • ✓ Evaluate imaging quality and tissue preservation with PFD as infiltrative medium
Protocol

Biopharma Insights We describe the use of perfluorodecalin as an infiltrative mounting medium. This is a simple method for improving depth of imaging in Arabidopsis thaliana leaf tissue with minimal physiological impact.

Difficulty
beginner
Total time
~15–20 min per leaf sample
Model organism
Arabidopsis thaliana

Steps

1
Mount Arabidopsis leaf samples in perfluorodecalin

Position freshly excised Arabidopsis leaves on a glass slide or chamber, then carefully infiltrate the leaf tissue with perfluorodecalin as the mounting medium to reduce light scattering and improve imaging depth penetration.

▶ 01:54
2
Image mounted leaf tissue using confocal microscopy

Acquire high-resolution fluorescence or confocal images of the PFD-infiltrated leaf samples, scanning through tissue depth to visualize cellular structures with enhanced optical clarity and minimal physiological disruption.

▶ 03:20
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