Protocol for detecting intracellular aggregations in Arabidopsis thaliana cell wall mutants using FM4-64 staining.

Abstract

Here, we present a step-by step protocol to visualize intracellular aggregations in Arabidopsis mutants with cell wall secretion defects using FM4-64, a lipophilic styryl dye. We describe steps for growing seedlings, staining them with FM4-64, and identifying intracellular aggregates in cell wall synthesis and/or secretion mutants in root and hypocotyl epidermal cells via confocal microscopy. Additionally, we provide troubleshooting suggestions for common pitfalls. For complete details on the use and execution of this protocol, please refer to Hoffmann and McFarlane. 1 .

Plant phenotyping relevance

Arabidopsis細胞内凝集体をFM4-64染色と共焦点顕微鏡で可視化・同定する実行手順とトラブルシューティングが中心であり、植物細胞状態の画像計測プロトコルに該当する。

Code and data availability

This is a methods protocol paper; the authors explicitly state 'This study did not generate unique datasets or code.' Fiji and NIS Elements are generic third-party software, not paper-specific assets. No public phenotype datasets, images, code, or models are deposited.

No evidence-backed public reproduction asset is currently recorded.