Abstract：
:In situ mRNA hybridization is one of the most powerful techniques for analyzing patterns of gene expression. However, its usefulness is limited in complex plant tissues by the need to fix, embed and section samples before localizing the desired mRNA. Here we present a robust whole-mount in situ hybridization method that allows easy access to patterns of gene expression in intact, complex tissues, such as the inflorescence apex of Arabidopsis thaliana. The tissue is first fixed and then permeabilized by treatment with a cocktail of cell wall-digesting enzymes that has been optimized to preserve the integrity of tissue structures, while also permitting the detection of expression patterns in deep tissues. In addition to colorimetric staining, fluorimetric staining that can be imaged by confocal microscopy can also be used with this protocol, thus providing full 3D resolution. The entire procedure can take <3 d from tissue preparation to image acquisition.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Rozier F,Mirabet V,Vernoux T,Das Pdoi
10.1038/nprot.2014.162subject
Has Abstractpub_date
2014-10-01 00:00:00pages
2464-75issue
10eissn
1754-2189issn
1750-2799pii
nprot.2014.162journal_volume
9pub_type
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