PUBLICATION

A Robust Procedure for Distinctively Visualizing Zebrafish Retinal Cell Nuclei under Bright Field Light Microscopy

Authors
Fu, J., Fang, W., Zou, J., Sun, M., Lathrop, K., Su, G., and Wei, X.
ID
ZDB-PUB-121206-40
Date
2013
Source
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society   61(3): 248-256 (Journal)
Registered Authors
Sun, Ming, Wei, Xiangyun
Keywords
feulgen nuclei staining, zebrafish, retina, photoreceptor, aging, degeneration
MeSH Terms
  • Animals
  • Azure Stains/analysis
  • Cell Nucleus/ultrastructure*
  • Coloring Agents/analysis*
  • Methylene Blue/analysis
  • Microscopy/methods
  • Neutral Red/analysis
  • Photoreceptor Cells, Vertebrate/ultrastructure*
  • Plastic Embedding/methods
  • Retina/cytology*
  • Retina/ultrastructure
  • Rosaniline Dyes/analysis
  • Staining and Labeling/methods*
  • Zebrafish/anatomy & histology*
PubMed
23204114 Full text @ J. Histochem. Cytochem.
Abstract

To simultaneously visualize individual cell nuclei and tissue morphologies of the zebrafish retina under bright field light microscopy, it is necessary to establish a procedure that specifically and sensitively stains the cell nuclei in thin tissue sections. This necessity arises from zebrafish retina’s high nuclear density and cone photoreceptors’ highly de-condensed chromatin, which significantly reduces their nuclear signals and makes nuclei difficult to distinguish from possible high cytoplasmic background staining. Here we optimized a procedure that integrates JB4 plastic embedding and Feulgen reaction for visualizing zebrafish retinal cell nuclei under bright field light microscopy. This method produces highly specific nuclear staining with minimal cytoplasmic background, allowing us to distinguish individual retinal nuclei despite the tight packaging of retinal nuclei. The nuclear staining is also sensitive enough to distinguish the euchromatin from heterochromatin in zebrafish cone nuclei. In addition, this method can be combined with in situ hybridization to simultaneously visualize the cell nuclei and mRNA expression patterns. With its superb specificity and sensitivity, this method may be extended to quantify cell density and analyze global chromatin organization throughout the retina or other tissues.

Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping