A layered mounting method for extended time-lapse confocal microscopy of whole zebrafish embryos

Sanat Upadhyay, Leoncio Vergara, Pranjali Shah, Jan Åke Gustafsson, Ioannis Kakadiaris, Maria Bondesson

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Dynamics of development can be followed by confocal time-lapse microscopy of live transgenic zebrafish embryos expressing fluorescence in specific tissues or cells. A difficulty with imaging whole embryo development is that zebrafish embryos grow substantially in length. When mounted as regularly done in 0.3-1% low melt agarose, the agarose imposes growth restriction, leading to distortions in the soft embryo body. Yet, to perform confocal time-lapse microscopy, the embryo must be immobilized. This article describes a layered mounting method for zebrafish embryos that restrict the motility of the embryos while allowing for the unrestricted growth. The mounting is performed in layers of agarose at different concentrations. To demonstrate the usability of this method, whole embryo vascular, neuronal and muscle development was imaged in transgenic fish for 55 consecutive hours. This mounting method can be used for easy, low-cost imaging of whole zebrafish embryos using inverted microscopes without requirements of molds or special equipment.

Original languageEnglish (US)
Article numbere60321
JournalJournal of Visualized Experiments
Volume2020
Issue number155
DOIs
StatePublished - Jan 2019

Keywords

  • Confocal microscopy
  • Developmental Biology
  • Embryo
  • Issue 155
  • Mounting
  • Time-lapse
  • Transgenic
  • Zebrafish

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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