Analysis of cellulose synthase activity in Arabidopsis using spinning disk microscopy

STAR Protoc. 2021 Oct 5;2(4):100863. doi: 10.1016/j.xpro.2021.100863. eCollection 2021 Dec 17.

Abstract

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.

Keywords: Cell Biology; Microscopy; Model Organisms; Plant sciences.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis Proteins / analysis*
  • Arabidopsis Proteins / chemistry
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism
  • Arabidopsis* / chemistry
  • Arabidopsis* / metabolism
  • Glucosyltransferases / analysis*
  • Glucosyltransferases / chemistry
  • Glucosyltransferases / genetics
  • Glucosyltransferases / metabolism
  • Hypocotyl* / chemistry
  • Hypocotyl* / metabolism
  • Microscopy / methods*

Substances

  • Arabidopsis Proteins
  • Glucosyltransferases
  • cellulose synthase