Fluorescence of TCS:GFP was observed in 8-day-old seedlings (T2) of wild types or abci20–1/21 expressing TCS:GFP using Olympus FLUOVIEW FV1000 confocal laser scanning microscope system. Excitation and emission were at 488 nm, and 520 nm, respectively. Fluorescent intensity was measured in the root tip and calculated using Image J (https://imagej.nih.gov/ij/index.html ; Schneider et al. 2012 (link)). The background fluorescence intensity was subtracted from the total intensity, following the method described previously (Ali et al. 2019 (link)).
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Quantifying GFP Fluorescence in Seedling Roots
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Other organizations : Pohang University of Science and Technology, University of California, San Diego, Nagoya University
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Variable analysis
independent variables
- Genotype (wild type or abci20-1/21 mutant)
- Fluorescence intensity of TCS:GFP in the root tip
- Age of seedlings (8-day-old)
- Excitation and emission wavelengths (488 nm and 520 nm, respectively)
- Imaging equipment (Olympus FLUOVIEW FV1000 confocal laser scanning microscope system)
- Image analysis software (ImageJ)
- Negative control: Background fluorescence intensity was subtracted from the total intensity
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