Immunocytochemistry as a tool for zebrafish developmental neurobiology

Authors: Novak A.E.¹; Ribera A.B.²

Source: Methods in Cell Science, Volume 25, Numbers 1-2, 2003 , pp. 79-83(5)

Publisher: Springer

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Abstract:

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.

Keywords: AEC (3-Amino-9-ethylcarbazole); Double in situ hybridization/immunocytochemistry; Fast Red; Immunocytochemistry; Zebrafish

Document Type: Research article

Affiliations: 1: 4200 E. 9th Ave, Department of Physiology and Biophysics C-240, University of Colorado Health Sciences Center, Denver, CO 80262, USA, Email: Alicia.novak@uchsc.edu 2: University of Colorado Health Sciences Center, Department of Physiology and Biophysics, Denver, CO, USA

Publication date: 2003-01-01