Stellaris smFISH probes targeting swt-3, an ortholog of the human solute carrier transporter SLC50A1, are shown in red (Cal Fluor 610), DAPI in blue, and PGL-1::GFP, a marker for P granules, in green. A) swt-3 transcripts are expressed in Z1/Z4 cells during embryogenesis (arrowheads). The embryo shows Z1 and Z4 migrating to Z2/Z3 P granule cells (marked with PGL-1::GFP in green). B) A dissected proximal gonad of a young adult capturing the ovulation of a recently fertilized zygote (arrow) as it passes through the swt-3 expressing spermatheca (red). Proximal oocyte marked with arrowhead. C) Sperm nuclei (blue) within the swt-3 (red) spermatheca. scale = 20µ The experiment confirms and extends swt-3 enrichment in cell-specific Z1/Z4 mRNA profiling experiment reported by Kroetz & Zarkower 2015.
The Stellaris acdh-1 smFISH probe contains 44 oligos labeled with Cal Fluor 610 (sequences available upon request). Staining was performed as described by Ji and van Oudenaarden (2012). In short, PGL-1::GFP embryos and worms were fixed with formaldehyde and ethanol. Hybridization was done in the dark at 37°C for four hours. Cal Fluor 610 coupled probes were designed with the Stellaris Probe Designer from Biosearch Technologies.
Reviewed ByAndy Golden
HistoryReceived: July 4, 2017
Accepted: July 6, 2017
Published: July 20, 2017