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Advances in Two-Photon Imaging in Plants
Yoko Mizuta1,2
1Institute for Advanced Research (IAR), Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi 464-8601, Japan.
Plant & Cell Physiology
|May 21, 2021
Summary
Two-photon excitation microscopy (2PEM) enables deep, high-resolution live imaging in plants by reducing autofluorescence. This advanced technique also allows for precise optical manipulation of plant cells and organelles.
Area of Science:
- Plant Science
- Microscopy
- Biotechnology
Background:
- Live and deep imaging are crucial for organismal physiology and biology.
- Two-photon excitation microscopy (2PEM) allows deep imaging of living tissues using less invasive near-infrared (NIR) lasers.
- 2PEM offers advantages over confocal microscopy, including reduced autofluorescence and deeper tissue penetration.
Purpose of the Study:
- To review recent advances in two-photon imaging.
- To discuss the applications of 2PEM in plant studies.
- To highlight the benefits of 2PEM for in vivo imaging in plant sciences.
Main Methods:
- Utilizing near-infrared (NIR) pulse lasers for two-photon excitation microscopy.
- Employing fluorescent proteins with long emission wavelengths (e.g., orange-red).
- Applying plant-specific clearing methods for enhanced imaging.
- Performing optical cell manipulations with femtosecond pulsed lasers.
Main Results:
- NIR 2PEM achieves greater depth and high-resolution imaging in plant tissues by reducing autofluorescence compared to confocal microscopy.
- Long-wavelength fluorescent proteins are suitable for two-photon live imaging in plants.
- Plant-specific clearing methods further improve deep and high-resolution imaging.
- Optical manipulation techniques enable single-cell or organelle level interventions.
Conclusions:
- Two-photon excitation microscopy is a powerful tool for advancing plant science research.
- 2PEM facilitates detailed studies of cellular communication and development in plants.
- Advances in 2PEM will significantly benefit in vivo imaging and biological studies in plants.
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