Rapid in-solution preparation of somatic and meiotic plant cell nuclei for high-quality 3D immunoFISH and immunoFISH-GISH.

ABSTRACT:

Background

Though multicolour labelling methods allow the routine detection of a wide range of fluorescent (immuno)probe types in molecular cytogenetics, combined applications for the simultaneous in situ detection of proteins and nucleic acids are still sporadic in plant cell biology. A major bottleneck has been the availability of high-quality plant nuclei with a balance between preservation of 3D ultrastructure and maintaining immunoreactivity. The aim of this study was to develop a quick and reliable procedure to prepare plant nuclei suitable for various combinations of immunolabelling and fluorescence in situ hybridisation methods (immunoFISH-GISH).

Results

The mechanical removal of the cell wall and cytoplasm, instead of enzymatic degradation, resulted in a gentle, yet

SUBMITTER: Makai D 

PROVIDER: S-EPMC10408160 | biostudies-literature | 2023 Aug

REPOSITORIES: biostudies-literature