RRC ID 60689
Author Saito S, Kawamura K, Matsuda Y, Suzuki T.
Title Brilliant Blue as an alternative dye to Fast Green for in ovo electroporation.
Journal Dev Growth Differ
Abstract 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.
Volume 61(7-8)
Pages 402-409
Published 2019-9
DOI 10.1111/dgd.12629
PMID 31612477
MeSH Animals Benzenesulfonates / chemistry* Chick Embryo Electroporation / methods* Fluorescence Gene Transfer Techniques Green Fluorescent Proteins / chemistry Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism* Luminescent Proteins / chemistry Luminescent Proteins / genetics Luminescent Proteins / metabolism* Microscopy, Fluorescence Reproducibility of Results Rosaniline Dyes / chemistry* Transgenes / genetics
IF 1.638
Times Cited 0
Resource
Chicken / Quail