ZebraFISH: Fluorescent In Situ Hybridization Protocol and Three-Dimensional Imaging of Gene Expression Patterns

被引：46

作者：

Welten, Monique C. M. ^{[1
,2
]}

De Haan, Simon B. ^{[1
,2
]}

van den Boogert, Niels ^{[1
]}

Noordermeer, Jasprien N. ^{[3
]}

Lamers, Gerda E. M. ^{[2
]}

Spaink, Herman P. ^{[2
]}

Meijer, Annemarie H. ^{[2
]}

Verbeek, Fons J. ^{[1
]}

机构：

[1] Leiden Univ, Leiden Inst Adv Comp Sci, NL-2333 CA Leiden, Netherlands

[2] Leiden Univ, Inst Biol, NL-2333 CA Leiden, Netherlands

[3] Leiden Univ, Med Ctr, Dept Mol Cell Biol, NL-2333 CA Leiden, Netherlands

来源：

ZEBRAFISH | 2006年 / 3卷 / 04期

关键词：

D O I：

10.1089/zeb.2006.3.465

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

We present a method and protocol for fluorescent in situ hybridization (FISH) in zebrafish embryos to enable three-dimensional imaging of patterns of gene expression using confocal laser scanning microscopy. We describe the development of our protocol and the processing workflow of the three-dimensional images from the confocal microscope. We refer to this protocol as zebraFISH. FISH is based on the use of tyramide signal amplification (TSA), which results in highly sensitive and very localized fluorescent staining. The zebraFISH protocol was extensively tested and here we present a panel of five probes for genes expressed in different tissues or single cells. FISH in combination with confocal laser scanning microscopy provides an excellent tool to generate three-dimensional images of patterns of gene expression. We propose that such three-dimensional images are suitable for building a repository of gene expression patterns, complementary to our previously published three-dimensional anatomical atlas of zebrafish development (bio-imaging.liacs.nl/). Our methodology for image processing of three-dimensional confocal images allows an analytical approach to the definition of gene expression domains based on the three-dimensional anatomical atlas.

引用

页码：465 / 476

页数：12

共 22 条

[11] Opinion - Cancer genetics and drug discovery in the zebrafish [J].

Stern, HM ;

Zon, LI .

NATURE REVIEWS CANCER, 2003, 3 (07) :533-539

[12]

Thisse B, 2004, METHOD CELL BIOL, V77, P505

[13]

THISSE C, 1993, DEVELOPMENT, V119, P1203

[14]

Verbeek FJ, 2004, PROC SPIE, V5304, P13

[15]

Verbeek FJ, 2002, PROC SPIE, V4672, P20

[16]

Verbeek FJ, 1999, INT J DEV BIOL, V43, P761

[17]

Verbeek FJ, 2000, PROC SPIE, V3964, P242

[18]

Verbeek FJ, 2000, IMAGE PROCESSING PRA, P153

[19]

Weinberg ES, 1996, DEVELOPMENT, V122, P271

[20]

Wilkinson DG, 1998, IN SITU HYBRIDIZATIO

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