Quantification of plant cell coupling with live-cell microscopy

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Movement of nutrients and signaling compounds from cell to cell is an essential process for plant growth and development. To understand processes such as carbon allocation, cell communication, and reaction to pathogen attack it is important to know a specific molecule’s capacity to pass a specific cell wall interface. Transport through plasmodesmata, the cell wall channels that directly connect plant cells, is regulated not only by a fixed size exclusion limit, but also by physiological and pathological adaptation. The noninvasive approach described here offers the possibility of precisely determining the plasmodesmata-mediated cell wall permeability for small molecules in living cells.

The method is based on photoactivation of the fluorescent tracer caged fluorescein. Non-fluorescent caged fluorescein is applied to a target tissue, where it is taken up passively into all cells. Imaged by confocal microscopy, loaded tracer is activated by UV illumination in a target cell and its spread to neighboring cells monitored. When combined with high-speed acquisition by resonant scanning or spinning disc confocal microscopy, the high signal-to-noise ratio of photoactivation allows collection of three-dimensional (3D) time series. These contain all necessary functional and anatomical data to measure cell coupling in complex tissues noninvasively.
Original languageEnglish
Title of host publicationPlasmodesmata : methods and protocols
EditorsManfred Heinlein
Number of pages12
VolumeIV
PublisherSpringer
Publication date2015
Pages137-148
Chapter9
ISBN (Print)978-1-4939-1522-4
ISBN (Electronic)978-1-4939-1523-1
Publication statusPublished - 2015
SeriesMethods in Molecular Biology
Volume1217
ISSN1064-3745

ID: 162904941