Isolation of Individual Cells and Tissues from Electroporated Sea Squirt (Ciona) Embryos by Fluorescence-Activated Cell Sorting (FACS)

Abstract

INTRODUCTIONThis protocol describes the dissociation of electroporated sea squirt (Ciona) embryos and the subsequent isolation of individual cells, cell lineages, and tissues, using fluorescence-activated cell sorting (FACS). Sorted samples can be used for RNA extraction and reverse transcriptase polymerase chain reaction (RT-PCR) or microarray analysis. Alternatively, dissociated samples can be used for the subsequent culturing and/or chemical treatment of isolated embryonic cells. This method permits the identification of effector genes responsible for changes in cell shape, polarity, or motility in a given tissue.