Optimization of Preparation and Detection Method for Pollen Tube Cell Wall by Atomic Force Microscopy
Abstract: Atomic force microscopy (AFM) is a powerful tool for the researches of the ultrastructure and mechanical properties of plant cell walls, while good sample preparation is essential for AFM data acquisition. As micron-scale plant samples, pollen tubes are typical experimental materials for studying the structure and function of cell walls. Due to the difficulty of sample preparation, it is hard to obtain in-situ AFM data in physiological state of pollen tubes as pollen tubes are hard to attach to the substrate firmly in fluid. In this study, the AFM preparation and detection method is optimized using Nicotiana tobacco pollen tubes as the experimental materials. Thin solid medium is used as the adhesive, thereby pollen tubes are attached to the glass slide during germination and elongation in fluidic environment. Compared with the conventional drying-rehydration method, the optimized liquid-attaching method allows observing directly in fluid without drying treatment, which avoids the shrinkage and denaturation of pollen tubes caused by drying-rehydration process. Accordingly, liquid-attaching method can be applied for pollen tubes of different species and sizes, providing support to get high-resolution in-situ AFM data of pollen tube cell walls in native physiological state under aqueous conditions.
Key words: atomic force microscopy, pollen tube, cell wall, in-situ, mechanical properties
CLC Number:
Q66
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