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Near-Infrared Autofluorescence Imaging of Nuclei in Living Plant Roots
Akira Yoshinari1,2, Masayoshi Nakamura1,2
1Institute for Advanced Research, Nagoya University, Nagoya, Japan.
Bio-Protocol
|April 28, 2025
Summary
Plant cell nuclei can be visualized using their natural near-infrared autofluorescence, derived from phytochrome. This less invasive live-cell imaging method works with conventional confocal microscopy for various plant tissues.
Area of Science:
- Plant Biology
- Cellular Imaging
- Biophotonics
Background:
- Autofluorescence in live-cell imaging typically interferes with target visualization.
- However, autofluorescence can serve as a useful marker for specific organelles.
- Phytochrome-derived autofluorescence from plant nuclei has been recently identified.
Purpose of the Study:
- To present protocols for visualizing near-infrared (NIR) nuclear autofluorescence in *Arabidopsis thaliana*.
- To enable less invasive live-cell imaging of plant nuclei.
- To facilitate fluorescence live-cell imaging in non-model plants.
Main Methods:
- Cultivating *Arabidopsis thaliana* plants under specific conditions.
- Utilizing confocal microscopy equipped with a 640 nm laser for imaging.
- Optimizing protocols for capturing NIR autofluorescence from root cells and pollen nuclei.
Main Results:
- Demonstrated successful visualization of plant nuclei via endogenous NIR autofluorescence.
- Established protocols for growing plants and performing confocal imaging of nuclear autofluorescence.
- Showcased the applicability of the method in root cells and pollen.
Conclusions:
- Phytochrome-derived NIR autofluorescence offers a novel, less invasive method for live-cell imaging of plant nuclei.
- This technique expands the possibilities for live-cell imaging in both model and non-model plant species.
- The presented protocols provide a foundation for further research into nuclear dynamics and function in plants.
