Automated whole mount localisation techniques for plant seedlings Journal Article


Author(s): Friml, Jiří; Benková, Eva; Mayer, Ulrike; Palme, Klaus; Muster, Gerhard
Article Title: Automated whole mount localisation techniques for plant seedlings
Affiliation
Abstract: Plant biology is currently experiencing a growing demand for easy and reliable mRNA and protein localisation techniques. Here, we present novel whole mount in situ hybridisation and immunolocalisation protocols, suitable to localise mRNAs and proteins in Arabidopsis seedlings. We demonstrate that these methods can be used in different organs of Arabidopsis seedlings as well as in other plant species. In order to achieve better reproducibility and higher throughput, we modified these protocols for automation to be performed by a liquid handling robot. In addition, we show that other procedures such as reporter enzyme assays and tissue clearing can be similarly automated. We present examples of application of our protocols including mRNA localisation and proteins and epitope tag (co)localisations which demonstrate that these methods provide reliable and versatile tools for expression, localisation and anatomical studies in plants.
Keywords: Epitope tag; Immunolocalisation; In situ hybridisation; InsituPro; Whole mount
Journal Title: Plant Journal
Volume: 34
Issue 1
ISSN: 0960-7412
Publisher: Wiley  
Date Published: 2003-04-01
Start Page: 115
End Page: 124
DOI: 10.1046/j.1365-313X.2003.01705.x
Open access: no