Brilliant Blue as an alternative dye to Fast Green for in ovo electroporation

• Published in: Development, growth & differentiation growth & differentiation, 2019-09, Vol.61 (7-8), p.402-409
• Main Authors: Saito, Seiji, Kawamura, Kazuki, Matsuda, Yoichi, Suzuki, Takayuki
• Format: Article
• Language: English
• Subjects: Animals; Benzenesulfonates - chemistry; Chick Embryo; Deoxyribonucleic acid; Developmental biology; DNA; Dyes; Electroporation; Electroporation - methods; Embryos; Fluorescence; fluorescent protein; Gene Transfer Techniques; Green Fluorescent Proteins - chemistry; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Injection; Luminescent Proteins - chemistry; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Microscopy, Fluorescence; quantification; Red Fluorescent Protein; Reproducibility of Results; Rosaniline Dyes - chemistry; Transgenes; Transgenes - genetics
• ISSN: 0012-1592; 1440-169X
• DOI: 10.1111/dgd.12629

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. We found Brilliant Blue as a new dye for the detection of red fluorescent proteins following chick embryo electroporation. We show useful examples for the use of Brilliant Blue for precise quantification of two fluorescence intensities after EGFP and mCherry co‐electroporation.