INIQUI   05448
INSTITUTO DE INVESTIGACIONES PARA LA INDUSTRIA QUIMICA
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Light sheet fluorescence microscopy for calcium dynamics in root and root hair cells of Arabidopsis thaliana
Autor/es:
COSTA, ALEX; CANDEO, ALESSIA; DOCCULA, FABRIZIO GANDOLFO; VALENTINI, GIANLUCA; ROMANO ARMADA, NELI; BASSI, ANDREA
Lugar:
Florencia
Reunión:
Congreso; Society for Experimental Biology Annual Metting, Masters of Biology; 2018
Institución organizadora:
Society for Experimental Biology
Resumen:
The accurate study of plant roots requires high-resolution microscopy, able to perform volumetric acquisitions in a few seconds, inducing as low photo-bleaching and photo-toxicity as possible to the living sample/organism. Light sheet fluorescence microscopy (LSFM) offers these characteristics, with the further advantage to mount the sample in a vertical position, mimicking the plant?s physiological growth conditions. Because of its transparency and its limited number of cells, the root of Arabidopsis thaliana represents a benchmark system to carry out imaging analyses. Root cells respond to several environmental or developmental stimuli by changing the intracellular free calcium concentration. These changes are commonly referred to as "calcium signatures" and can range from a single transient increase to a series of repetitive calcium oscillations. In the talk, I will report the application of LSFM for calcium imaging in root tip cells of Arabidopsis plants expressing the intensimetric- (GCaMP3) or the Förster Resonance Energy Transfer (FRET)-based (Cameleon YC3.6) calcium sensors. I will show that the fast acquisition rate of LSFM enables both 3D (x,y,t) and 4D (x,y,z,t) imaging analyses. Particularly, I will present that the 4D imaging reveals the existence of a calcium signal percolation phenomenon inside the root body. Finally, I will show that LSFM is ideally suited to image several growing root hairs simultaneously and to follow their oscillatory tip-focused calcium gradients, correlating them with the rate of hairs growth.