Isolation of <i>C. elegans</i> neurons from larval animals by Fluorescence Activated Cell Sorting (FACS).

<p>(A) Expression of the pan-neural GFP marker, <i>evIs111</i>, in an L2 stage larval animal. Anterior is to left. (B) Procedure for generating dissociated suspensions of larval cells for primary culture and for isolation of GFP-marked cells (green) by FACS. (C) Primary culture of L1-stage cells 36 hr after plating. Arrows point to elongated processes extending from GFP-labeled neurons. Scatterplot of FACS profile for cells dissociated from the wild-type (N2) reference strain (D) and from the pan-neural <i>evIs111</i> line (E). Propidium Iodide (PI) marks dead cells (blue). GFP-labeled cells (green) were isolated by FACS (outlined with box). The majority (∼90%) of FACS-derived cells (F) are marked with GFP (G) in primary cultures examined within 2 hr of FACS isolation. Note that neurons are loosely attached which likely accounts for the displacement of DIC vs GFP images of individual cells in these micrographs. Scale bars = 10 micron.