Fast and High-Resolution Imaging of Pollinated Stigmatic Cells by Tabletop Scanning Electron Microscopy.

Area of Science:

• Plant reproductive biology
• Microscopy techniques
• Cellular imaging

Background:

• Successful plant reproduction relies on the critical initial interaction between pollen and stigma epidermal cells.
• Pollen germination and subsequent pollen tube growth are essential for transporting sperm cells to ovules.
• Confocal microscopy, while useful, is time-consuming and requires fluorescent marker lines for studying post-pollination events.

Purpose of the Study:

• To propose a quick, high-resolution imaging protocol for analyzing pollen-stigma interactions.
• To overcome the limitations of time-consuming methods like confocal microscopy.
• To provide an efficient way to track pollen tube growth and behavior within the stigma.

Main Methods:

• Utilized tabletop scanning electron microscopy (SEM) for imaging.
• Developed a protocol that does not require prior sample fixation or fluorescent marker lines.
• Captured images from early pollen hydration (minutes post-pollination) to pollen tube growth (1 hour post-pollination).

Main Results:

• The tabletop SEM protocol provides high-resolution images of pollen grain behavior on stigmatic cells.
• Effectively captures early pollen hydration and subsequent pollen tube growth within the stigma.
• Demonstrated efficiency in tracking pollen tube paths, applicable to *Arabidopsis thaliana* and potentially other species.

Conclusions:

• Tabletop SEM offers a rapid and efficient alternative for studying pollen-stigma interactions.
• This method facilitates the analysis of pollen hydration states and pollen tube phenotypes across different genetic backgrounds.
• The protocol is broadly applicable for research in plant reproductive biology and genetics.