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Fig. 3 Figure 3. Additives improve signal detection. Nitro-blue tetrazolium chloride paired with 5-bromo-4-chloro-3-indolyl phosphate (NBT/BCIP) staining of atoh1b compared with and without volume excluding additives in 24 h postfertilization zebrafish embryos. (A–F) Embryos from the same batch were stained until first color appearance in controls (A & D). (G–L) Embryos stained to completion. (B, E, H & K) Polyvinyl alcohol (10%) enhances signal intensity of real and background stain (eye region; arrowheads). (C, F, I & L) Dextran sulfate (5%) increases sensitivity (hindbrain; arrow) without increasing background. Embryos were manually deyolked. Scale bar = 100 μm.