Simultaneous quantitative detection of different mRNAs in whole embryos.
cxcl12a expression pattern (A) and vasa transcript (B) in PGCs are specifically detected in 12-hpf zebrafish embryos. myoD RNA expression can be detected in the myotomes of the same sample (C) and cell nuclei are stained with DAPI (D). (E) An overlay of all the channels. (F) 12-hpf zebrafish embryo expressing cxcl12a and vasa transcripts. The left side of the embryo has the cxcl12a expression pattern in pseudocolors together with PGCs in magenta, demonstrating the arrival of the PGCs at domains of high cxcl12a expression. The calibration bar is shown for the left half of the embryo. The right side of the embryo shows cxcl12a expression domains in green, with the PGCs expressing vasa in red and nuclei stained in blue. The intensity profile of cxcl12a expression along the white line is presented in the graph on the right in arbitrary units (a.u.). Confocal images of whole-mount zebrafish embryos were captured using a 20× objective and 0.6× digital zoom in 3-μm intervals and subsequently processed as z-projections using the ImageJ software.