Fig. 1

Dual-color VAEM fluorescence co-localization with organelle markers is ideal for dissecting protein localization and dynamics in the cell cortex, taking Auxin Binding Protein 1 (ABP1) as an example. The punctate structures in VAEM images were tracked by using spatially and temporally global particle assignment with MATLAB. The incident angles were between 63.03° and 66.64° for HDEL-GFP seedlings, corresponding to penetration depths at 120 nm (66.64°) – 250-300 nm (63.03°) respectively, depending on the practical conditions for the adherence of seedling. a Transgenic Arabidopsis line expressing ABP1:ABP1-YFP (ABP1:ABP1-YFP / Col-0) showing subcellular localization to plasma membrane and intracellular structures, circles indicates punctate structures tagged by YFP fluorescence; b Transgenic Arabidopsis line expressing ABP1:ABP1-YFP showing blurred fluorescence, corresponding bright field images were shown in the inlets; c Transgenic Arabidopsis line expressing ABP1:ABP1-YFP showing uniformed distribution to ER and punctate structures under VAEM; d The punctate structures partly co-localize to Golgi apparatuses but do not co-localize to mitochondria; e Some ABP1-YFP tagged punctate structures frequently co-diffuse with Golgi apparatuses. The time series are shown for every 4 images. Bars = 200 μm (a), 7 μm (b-d), 5 μm (e). Interval between frames = 200 ms