Immunostaining of dissected zebrafish embryonic heart

Jingchun Yang, Xiaolei H Xu

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Zebrafish embryo becomes a popular in vivo vertebrate model for studying cardiac development and human heart diseases due to its advantageous embryology and genetics. About 100-200 embryos are readily available every week from a single pair of adult fish. The transparent embryos that develop ex utero make them ideal for assessing cardiac defects. The expression of any gene can be manipulated via morpholino technology or RNA injection. Moreover, forward genetic screens have already generated a list of mutants that affect different perspectives of cardiogenesis. Whole mount immunostaining is an important technique in this animal model to reveal the expression pattern of the targeted protein to a particular tissue. However, high resolution images that can reveal cellular or subcellular structures have been difficult, mainly due to the physical location of the heart and the poor penetration of the antibodies. Here, we present a method to address these bottlenecks by dissecting heart first and then conducting the staining process on the surface of a microscope slide. To prevent the loss of small heart samples and to facilitate solution handling, we restricted the heart samples within a circle on the surface of the microscope slides drawn by an immEdge pen. After the staining, the fluorescence signals can be directly observed by a compound microscope. Our new method significantly improves the penetration for antibodies, since a heart from an embryonic fish only consists of few cell layers. High quality images from intact hearts can be obtained within a much reduced procession time for zebrafish embryos aged from day 2 to day 6. Our method can be potentially extended to stain other organs dissected from either zebrafish or other small animals.

Original languageEnglish (US)
Article numbere3510
Pages (from-to)1-3
Number of pages3
JournalJournal of Visualized Experiments
Issue number59
DOIs
StatePublished - 2012

Fingerprint

Zebrafish
Embryonic Structures
Microscopes
Antibodies
Fishes
Fish
Animals
Staining and Labeling
Morpholinos
Embryology
Human Development
Image resolution
Vertebrates
RNA
Heart Diseases
Coloring Agents
Image quality
Animal Models
Fluorescence
Technology

Keywords

  • Cardiology
  • Danio rerio
  • Developmental biology
  • Dissection
  • Embryonic heart
  • Immunostaining
  • Issue 59
  • Zebrafish

ASJC Scopus subject areas

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

Cite this

Immunostaining of dissected zebrafish embryonic heart. / Yang, Jingchun; Xu, Xiaolei H.

In: Journal of Visualized Experiments, No. 59, e3510, 2012, p. 1-3.

Research output: Contribution to journalArticle

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