Growth conditions in vitro:

. 48hours at 4°C in the dark -> 4 days under 16hr/8hr (light/dark) light condition
. growth media: 1MS; 1% sucrose; 0.5g/l MES pH5.7; 0.8% agar

Genotypes:

. GFP-MBD (Marc et al., 1998) DOI: 10.1105/tpc.10.11.1927
. mCit-MBD (Armezzani et al., 2018) DOI: 10.1242/dev.162255
. GFP-TUA6 (Ueda et al., 1999) https://doi.org/10.1007/BF01279267
. bot1-7 / GFP-MBD (Uyttewaal et al., 2012) DOI: 10.1016/j.cell.2012.02.048

Imaging:

. Microscope: Zeiss LSM800 upright; Zen blue software 
. Objective: W Plan-Apochromat 40x/1.0 NA; water dipping
. Z-stacks: 0.5um Z steps
. Frame size: 1024x512 px (pixel size: 0.31um)
. Time series: 20min time interval during 4 hours

Folder architecture and name of the Tiff files:

The zip folder contains 4 folders for each genotype tested have their own folder. These folders contain each two subfolders, “ablation_Tiff” and “control_Tiff” containing the tif images of the experiments with or without ablation respectively. 
.tif images are named as follow: “genotype_samplenumber_timepoint.tif”. 

Related ressources:

. Zenodo: - https://zenodo.org/record/7436075#.Y5rmd-zMJF8 : contains all intermediate data generated by the image analysis (processed images, Regions of interest, quantifications, logs,...) from the raw .tif images available in the zip folder. 
          - https://zenodo.org/record/7434536#.Y7wJI_7MKgY (also available on GitHub: https://github.com/VergerLab/MT_Angle2Ablation_Workflow): contains all the scripts for the image analysis (imageJ macros and python script and a step by step description of the method) used on the raw .tif images available in the zip folder.