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Cell Biology JoVE (Open Access) Citable · DOI

Light Sheet Fluorescence Microscopy of Plant Roots Growing on the Surface of a Gel

DOI: 10.3791/55044-v
What you'll learn
  • ✓ Prepare Arabidopsis seedlings for light-sheet microscopy imaging
  • ✓ Mount plant samples vertically on gel surface for growth observation
  • ✓ Configure light-sheet microscope for long-term root development imaging
  • ✓ Acquire time-lapse fluorescence data of developing plant organs
Protocol

This protocol shows a plant sample preparation method for light-sheet microscopy. The setup is characterized by mounting the plant vertically on the surface of a gel and letting it grow in controlled bright conditions. This allows long-term observation of plant organ development in standardized conditions.

Difficulty
intermediate
Total time
~1 week (seedling growth) + ~30 min (sample prep and imaging setup)
Model organism
Arabidopsis thaliana

Steps

1
Cultivate Arabidopsis thaliana seedlings

Grow Arabidopsis seedlings under controlled light conditions until ready for mounting. Seedlings should be at an appropriate developmental stage for root imaging.

▶ 01:24
2
Prepare and mount plant sample on gel

Position the seedling vertically on the surface of a gel medium, securing it for stable imaging. The gel substrate supports growth while allowing optical access for light-sheet microscopy.

▶ 02:04
3
Configure light-sheet microscope for imaging

Align the light-sheet optics and fluorescence detection path. Set acquisition parameters for long-term time-lapse imaging of root development in standardized environmental conditions.

▶ 03:28
4
Acquire and visualize time-lapse image data

Record sequential fluorescence images over time to visualize plant organ development. Process and display results showing growth dynamics at cellular resolution.

▶ 05:18
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