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Analysis of cellulose synthase activity in Arabidopsis using spinning disk microscopy

ABSTRACT: Summary We describe sample preparation and visualization of fluorescently tagged cellulose synthases in cellulose synthase complexes at the plasma membrane of Arabidopsis hypocotyl epidermal cells using live-cell imaging via spinning disk microscopy. We present a technique for sample mounting that may be suitable for imaging other samples. Additionally, we offer free, open-source solutions for image analysis and provide extensive troubleshooting suggestions. For complete information on the use and execution of this protocol, please refer to McFarlane et al., 2021. Graphical abstract Highlights • Protocol to track cellulose synthesis in real time in living plant cells• Image analysis pipeline to quantify cellulose synthase complex speeds• Example dataset for analysis and example data display suggestions We describe sample preparation and visualization of fluorescently tagged cellulose synthases in cellulose synthase complexes at the plasma membrane of Arabidopsis hypocotyl epidermal cells using live-cell imaging via spinning disk microscopy. We present a technique for sample mounting that may be suitable for imaging other samples. Additionally, we offer free, open-source solutions for image analysis and provide extensive troubleshooting suggestions.

SUBMITTER: Verbancic J

PROVIDER: S-EPMC8503582 | biostudies-literature |

REPOSITORIES: biostudies-literature

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