Whole mount in situ method

Malcolm Bennett, University of Warwick
e-mail: bt@dna.bio.warwick.ac.uk

The whole mount in situ method is derived from the Tautz and Pfeifle (1989) paper (Chromosoma 98:81-85) which described the non-radioactive localisation of hunchback mRNA in Drosophila embryos. Ludevid et al (1992) have adapted the method in a Plant Physiol. paper (100, 1633-1639) to suit young Arabidopsis seedlings. Other groups, such as Kwart et al (1993) (Plant J., 4, 993-1002), have reported its use in localising differential gene expression. I would still recommend performing in situs using sections as well, but this provides a useful means to have a quick 'look and see' at your mRNAs spatial expression pattern.

The Ludevid et al (1992) paper provides only brief details of their modification to the Tautz and Pfeifle paper, so I've written out details of my protocol. All washing steps are conducted at room temperature (unless stated in the text) and samples are attached to a rotating wheel to facilitate washing. All solutions are DEPC-treated (except Tris containing ones) in case riboprobes are used.