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+## Comparison of autophagic activity in roots mounted on glass or in RoPods
+
+To assess the impact of glass-based sample mounting on the autophagic activity, RoPod v24 and standard agar plates were prepared similarly to the experiment for the root length measurement. After 6 days, seedlings growing on top of the agar medium were transferred in an empty glass bottom chamber, on a drop of 0.5 × MS and then gently covered with a cover slide. To prevent fast evaporation of the liquid medium, the glass bottom chamber was covered with a lid. Both RoPod v24 with seedlings and glass bottom chamber with transferred seedlings were imaged at different end points using CLSM 800 (Carl Zeiss), Objective 40x/1.2W, excitation light 488 nm, and emission 505–560 nm. The number of puncta was quantified in *Fiji* using the workflow presented in Supplementary File S2.
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