Sample Preparation for Fluorescence Imaging of the Cytoskeleton in Fixed and Living Plant Roots

By jpeza - Posted on 29 Noviembre 2010

Fecha Publicación: 
2 Oct 2009
Nombre de Revista: 
Datos del paper
Autor Principal: 
Julia Dyachok
Página Inicial: 
Página Final: 

During the past decade 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 root
cells, 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. Parallel to our live cell imaging
approaches, we routinely process fixed plant material via indirect
immunofluorescence. For these methods we typically use
nonembedded 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.

Dirección del Autor: 

Plant Biology Division, The Samuel Roberts Noble Foundation Inc., Ardmore, OK, USA

Actin - Arabidopsis - Microtubules - Green fluorescent protein - Living cells - Roots - Immunofluorescence - Fixed pl

Cheol-Min Yoo, Karuppaiah Palanichelvam,
Elison B. Blancaflor

[file] Methods_in_Molecular_Biology_2009_vol586_pp_157-169.pdf2.76 MB