Fluorescence imaging of the cytoskeleton in plant roots

Julia Dyachok, Ana Paez-Garcia, Cheol Min Yoo, Karuppaiah Palanichelvam, Elison B. Blancaflor

Research output: Chapter in Book/Report/Conference proceedingChapter

7 Scopus citations

Abstract

During the past two decades the use of live cytoskeletal probes has increased dramatically due to the introduction of the green fluorescent protein. However, to make full use of these live cell reporters it is necessary to implement simple methods to maintain plant specimens in optimal growing conditions during imaging. To image the cytoskeleton in living Arabidopsis roots, we rely on a system involving coverslips coated with nutrient supplemented agar where the seeds are directly germinated. This coverslip system can be conveniently transferred to the stage of a confocal microscope with minimal disturbance to the growth of the seedling. For roots with a larger diameter such as Medicago truncatula, seeds are first germinated in moist paper, grown vertically in between plastic trays, and roots mounted on glass slides for confocal imag- ing. Parallel with our live cell imaging approaches, we routinely process fixed plant material via indirect immunofluorescence. For these methods we typically use non-embedded vibratome-sectioned and whole mount permeabilized root tissue. The clearly defined developmental regions of the root provide us with an elegant system to further understand the cytoskeletal basis of plant development.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages139-153
Number of pages15
Volume1365
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1365
ISSN (Print)10643745

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Keywords

  • Actin
  • Arabidopsis
  • Fixed plant material
  • Green fluorescent protein
  • Immunofluorescence
  • Living cells
  • Medicago truncatula
  • Microtubules
  • Roots
  • Sectioning

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Dyachok, J., Paez-Garcia, A., Yoo, C. M., Palanichelvam, K., & Blancaflor, E. B. (2016). Fluorescence imaging of the cytoskeleton in plant roots. In Methods in Molecular Biology (Vol. 1365, pp. 139-153). (Methods in Molecular Biology; Vol. 1365). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3124-8_7