Immunolocalisation and in situ hybridisation allow the detection of proteins and RNA respectively, in individual cells of different tissue types. Hybridisation on serial 5–8 μm sections is most... Show moreImmunolocalisation and in situ hybridisation allow the detection of proteins and RNA respectively, in individual cells of different tissue types. Hybridisation on serial 5–8 μm sections is most frequently performed with tissue embedded in paraffin wax but it is often difficult to obtain high-resolution sections from soft tissue with this embedding process. To overcome this, alternative localisation protocols have been developed utilising plastic resins. We have used plastic embedded tissue from Lotus japonicus roots and young nodules successfully in immunolocalisation experiments and developed a protocol that can also be adapted for in situ RNA localisation studies. The different parameters tested are described, as well as the use of alkaline phosphatase- or fluorescently-conjugated secondary antibodies. Show less
