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PLoS One. 2013 Oct 16;8(10):e75646. doi: 10.1371/journal.pone.0075646. eCollection 2013.

Calcium dynamics in root cells of Arabidopsis thaliana visualized with selective plane illumination microscopy.

Author information

1
Dipartimento di Bioscienze, Università degli studi di Milano, Milano, Italy.

Abstract

Selective Plane Illumination Microscopy (SPIM) is an imaging technique particularly suited for long term in-vivo analysis of transparent specimens, able to visualize small organs or entire organisms, at cellular and eventually even subcellular resolution. Here we report the application of SPIM in Calcium imaging based on Förster Resonance Energy Transfer (FRET). Transgenic Arabidopsis plants expressing the genetically encoded-FRET-based Ca(2+) probe Cameleon, in the cytosol or nucleus, were used to demonstrate that SPIM enables ratiometric fluorescence imaging at high spatial and temporal resolution, both at tissue and single cell level. The SPIM-FRET technique enabled us to follow nuclear and cytosolic Ca(2+) dynamics in Arabidopsis root tip cells, deep inside the organ, in response to different stimuli. A relevant physiological phenomenon, namely Ca(2+) signal percolation, predicted in previous studies, has been directly visualized.

PMID:
24146766
PMCID:
PMC3797704
DOI:
10.1371/journal.pone.0075646
[Indexed for MEDLINE]
Free PMC Article

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