Accedi

Fondazione Filarete

2 ARTICLES PUBLISHED IN JoVE

Biology

Visualization of Endoplasmic Reticulum Subdomains in Cultured Cells

Matteo Fossati^1,2,3, Nica Borgese^2,3,4, Sara Francesca Colombo^2,3, Maura Francolini^1,2

¹Fondazione Filarete, ²Department of Biotechnology and Translational Medicine, University of Milan, ³Neuroscience Institute, National Research Council (CNR), ⁴Department of Health Science, "Magna Graecia" University of Catanzaro

We describe the imaging approaches we use to investigate the distribution and mobility of the transfected fluorescent proteins resident in the endoplasmic reticulum (ER) by means of the confocal imaging of living cells. We also ultrastructurally analyze the effect of their expression on the architecture of this subcellular compartment.

JoVE Core

Biosensing Motor Neuron Membrane Potential in Live Zebrafish Embryos

Lorena Benedetti^1,2,3, Anna Ghilardi⁴, Laura Prosperi⁴, Maura Francolini^*1, Luca Del Giacco^*4

¹Department of Medical Biotechnology and Translational Medicine, Università degli Studi di Milano, ²Department of Neuroscience; Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine, ³Program in Cellular Neuroscience, Neurodegeneration and Repair, Yale University School of Medicine, ⁴Department of BioSciences, Università degli Studi di Milano

Protocols described here allow for the study of the electrical properties of excitable cells in the most non-invasive physiological conditions by employing zebrafish embryos in an in vivo system together with a fluorescence resonance energy transfer (FRET)-based genetically encoded voltage indicator (GEVI) selectively expressed in the cell type of interest.