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ZFIN ID: ZDB-PUB-031229-12
Immunocytochemistry as a tool for zebrafish developmental neurobiology
Novak, A.E. and Ribera, A.B.
Date: 2003
Source: Methods in cell science : an official journal of the Society for In Vitro Biology   25(1-2): 79-83 (Review)
Registered Authors: Novak, Alicia, Ribera, Angie
Keywords: AEC (3-Amino-9-ethylcarbazole), Double in situ hybridization/immunocytochemistry, Fast Red, Immunocytochemistry, Zebrafish
MeSH Terms:
  • Animals
  • Embryo, Nonmammalian/metabolism
  • Fluorescent Dyes/chemistry
  • Immunoenzyme Techniques
  • Immunohistochemistry/methods*
  • In Situ Hybridization, Fluorescence
  • Neurons/metabolism*
  • Spinal Cord/metabolism*
  • Zebrafish/embryology
  • Zebrafish/genetics
  • Zebrafish/metabolism*
PubMed: 14739591 Full text @ Methods Cell Sci.
Two methods are presented here that allow clear visualization of antibody localization in zebrafish whole mount preparations, both for immunocytochemistry (ICC) alone and in combination with in situ hybridization (ISH). The first protocol describes ICC performed using a modified permeabilization technique and the chromogen AEC (3-Amino-9-ethylcarbazole). The second protocol describes the co-localization of transcriptional and translational products using a combined ISH/ICC protocol. A fluorescing chromogen (Fast Red, FR) is used to detect mRNA transcripts by ISH, and is combined with ICC that uses a secondary antibody conjugated to a different fluorescent molecule (Alexa 488). These procedures allow the identification of gene expression patterns in cell types identifiable with known antibodies.