DII images in root tips over time

Published: 16 May 2023| Version 1 | DOI: 10.17632/p7ftp5wm3h.1
, Anthony Bishopp


Data comprises timecourse for DII (DEAL) expression for 9 Arabidopsis roots. The raw data is deposited in .lif (Leica) format. This can be opened in ImageJ using the Bioformats plugin. Each file represents data from a single root. Each image within the file represents a timepoint from 0-18h. Each image represents a small z-stack (3 slices) taken from the centre of the root paired with images + 5um and - 5um to this.


Steps to reproduce

plant growth and preparation DII:Venus seeds cloned using the DEAL vector were sown on 0.5x Murishage and Skoog basal salts with 1% MES monohydrate, 1% agar, pH 5.7. Seeds were then stratified at 4°C in darkness for 2 days before being placed in a growth chamber at 21°C with a 16h light, 8h dark regime. 3-4 days after germination and 12 hours prior to imaging, seedlings were transferred to glass-bottomed imaging cuvettes and the root covered with a slice of growth medium gel. They were then returned to their growth conditions overnight. Imaging and image analysis Being kept in growth chambers in between imaging sessions, each seedling was imaged once per hour for 18 hours using a Leica SP8 laser scanning confocal microscope with a 40x dry objective. Excitation of both VENUS and tdTomato was performed using a 514 nm laser, and emission collected with photomultiplier tube or hybrid detectors across 520-550 nm and 570-760 nm, respectively.


University of Nottingham


Confocal Microscopy, Arabidopsis thaliana, Root, Auxin