Chick embryo electroporation is a powerful tool for the introduction of transgenes into tissues of interest for the study of developmental biology. This method often uses Fast Green to visualize the injected area by staining the solution containing DNA green. Here, we show that Fast Green fluoresces in a red color after electroporation, suggesting that researchers need to be cautious when detecting red fluorescence. Fast Green solution did not show any fluorescence before injection into chick embryos, but fluoresced red within 3 min post-injection into chick embryos. We identified Brilliant Blue as suitable alternative dye for use as an indicator of injection sites in ovo electroporation. We found that 0.2% of Brilliant Blue was sufficient to track the area of DNA injection. In addition, this chemical did not show red fluorescence after electroporation. Our findings demonstrate that Brilliant Blue can be used for detecting red fluorescent proteins introduced into chick embryos by electroporation. Our study also shows useful examples for the application of Brilliant Blue for the precise quantification of two fluorescence intensities after EGFP and mCherry co-electroporation.

中文翻译：

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灿烂的蓝色作为卵黄电穿孔中坚牢的绿色的替代染料。

小鸡胚胎电穿孔是将转基因引入感兴趣的组织以研究发育生物学的强大工具。该方法通常使用坚牢绿色，通过将包含DNA的溶液染成绿色来可视化注入的区域。在这里，我们显示了电穿孔后，坚牢的绿色发出红色荧光，这表明研究人员在检测红色荧光时需要谨慎。坚牢的绿色溶液在注射到鸡胚之前没有显示任何荧光，但在注射到鸡胚后3分钟内发出红色荧光。我们确定了亮蓝是适合用作卵电穿孔中注射部位指示剂的替代染料。我们发现0.2％的亮蓝足以追踪DNA注入的面积。此外，该化学试剂在电穿孔后没有显示红色荧光。我们的发现表明，亮蓝可用于检测通过电穿孔引入鸡胚的红色荧光蛋白。我们的研究还显示了应用亮蓝对EGFP和mCherry共电穿孔后两种荧光强度进行精确定量的有用实例。