Visualizing morphogenesis in transgenic zebrafish embryos using BODIPY TR methyl ester dye as a vital counterstain for GFP

• Published in: Developmental dynamics Vol. 232; no. 2; pp. 359 - 368
• Main Authors: Cooper, Mark S., Szeto, Daniel P., Sommers‐Herivel, Greg, Topczewski, Jacek, Solnica‐Krezel, Lila, Kang, Hee‐Chol, Johnson, Iain, Kimelman, David
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.02.2005
• Subjects: Animals; Animals, Genetically Modified; BODIPY TR dye; Boron Compounds - pharmacology; Coloring Agents - pharmacology; confocal microscopy; Danio rerio; Esters - pharmacology; Genetic Techniques; germ layers; green fluorescent protein; Green Fluorescent Proteins - chemistry; Green Fluorescent Proteins - pharmacology; Luminescent Proteins - chemistry; mesoscopic; Microscopy, Confocal - methods; Microscopy, Fluorescence; organogenesis; Software; Spectrophotometry; stem cells; Time Factors; time‐lapse; Transgenes; transgenic zebrafish; visualization; vital imaging; vital staining; Zebrafish
• ISSN: 1058-8388; 1097-0177
• DOI: 10.1002/dvdy.20252

Green fluorescent protein (GFP) technology is rapidly advancing the study of morphogenesis, by allowing researchers to specifically focus on a subset of labeled cells within the living embryo. However, when imaging GFP‐labeled cells using confocal microscopy, it is often essential to simultaneously visualize all of the cells in the embryo using dual‐channel fluorescence to provide an embryological context for the cells expressing GFP. Although various counterstains are available, part of their fluorescence overlaps with the GFP emission spectra, making it difficult to clearly identify the cells expressing GFP. In this study, we report that a new fluorophore, BODIPY TR methyl ester dye, serves as a versatile vital counterstain for visualizing the cellular dynamics of morphogenesis within living GFP transgenic zebrafish embryos. The fluorescence of this photostable synthetic dye is spectrally separate from GFP fluorescence, allowing dual‐channel, three‐dimensional (3D) and four‐dimensional (4D) confocal image data sets of living specimens to be easily acquired. These image data sets can be rendered subsequently into uniquely informative 3D and 4D visualizations using computer‐assisted visualization software. We discuss a variety of immediate and potential applications of BODIPY TR methyl ester dye as a vital visualization counterstain for GFP in transgenic zebrafish embryos. Developmental Dynamics 232:359–368, 2005. © 2004 Wiley‐Liss, Inc.