Expression of genes mfap4, cxcr3.2, mpeg1, and ptpn6 colocalizes with expression of macrophage-specific marker csf1r. Double fluorescent in situ with the csf1r gene. Probes for mRNA are used as indicated in the bottom left corner of each image. Expression of genes, mfap4 (A-C), cxcr3.2 (D-F), mpeg1 (G-I), and ptpn6 (J-L) overlaps with expression of macrophage-specific marker csf1r. Images show summed Z-stacks of consecutive confocal images through the posterior blood island of 28-hpf embryos (lateral view). A Zeiss axioplan microscope with Bio-Rad MRC1024ES scanhead was used for confocal imaging. Signals in the green and red channels were scanned sequentially using, respectively, the 488-nm laser line with 522 DF32 filter for detection of emitted light, and the 568-nm laser line with 605 DF32 filter for detection of emitted light: 10×/0.30 NA and 20×/0.50 NA Plan-Neofluar objectives were used. Images were processed in ImageJ Version 1.43 software and show (from left to right) the signal in the green channel, the signal in the red channel, and the merged signals. (A-F) Bars represent 20 μm. (G-L) Bars represent 60 μm.