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

Mark S. Cooper*, Daniel P. Szeto, Greg Sommers-Herivel, Jacek Topczewski, Lila Solnica-Krezel, Hee Chol Kang, Iain Johnson, David Kimelman

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

92 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)359-368
Number of pages10
JournalDevelopmental Dynamics
Volume232
Issue number2
DOIs
StatePublished - Feb 2005

Keywords

  • BODIPY TR dye
  • Confocal microscopy
  • Germ layers
  • Green fluorescent protein
  • Mesoscopic
  • Organogenesis
  • Stem cells
  • Time-lapse
  • Transgenic zebrafish
  • Visualization
  • Vital imaging
  • Vital staining

ASJC Scopus subject areas

  • Developmental Biology

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