Protocol: an improved and universal procedure for whole-mount immunolocalization in plants

Area of Science:

• Plant cell biology
• Microscopy
• Molecular biology

Background:

• Microscopy advances drive cell biology research.
• Reproducible tissue preservation and cell labeling for immunolocalization remain challenging.
• Analyzing inner cell layers, tissues, and organs requires robust protocols.

Purpose of the Study:

• To describe an improved, robust whole-mount immunolocalization protocol for diverse plant species.
• To enable in-depth microscopic analysis of protein localization and gene expression.
• To provide a time-efficient method without robotic equipment.

Main Methods:

• Optimized sample fixation and tissue clearing.
• Multi-protein staining compatible with nucleic acid detection.
• Optional cell wall and nucleus labeling for morphology and gene expression analysis.

Main Results:

• The protocol is applicable to a wide range of plant species.
• Achieves accurate, high-resolution, and reproducible protein detection.
• Allows simultaneous analysis of protein localization and gene expression patterns at single-cell resolution.

Conclusions:

• The developed protocol addresses key challenges in plant immunolocalization.
• Facilitates detailed morphological and gene expression studies with high precision.
• Offers a rapid, single-day solution for advanced plant cell imaging.