Multicolour three dimensional structured illumination microscopy of immunolabeled plant microtubules and associated proteins.

摘要: In the present work, we provide an account of structured illumination microscopy (SIM) imaging fixed and immunolabeled plant probes. We take advantage SIM, to superresolve intracellular structures at a considerable z-range circumvent its low temporal resolution capacity during study living samples. Further, validate protocol for transgenic material expressing fluorescent protein-based markers different subcellular structures. Focus is given on 3D bulky structures, such as mitotic cytokinetic microtubule arrays well performance SIM using multichannel quantitative correlations that can be deduced. As proof concept, superresolution output organization cortical microtubules in wild-type mutant Arabidopsis cells, including aberrant preprophase bands phragmoplasts cytoskeletal devoid p60 subunit severing protein KATANIN refined details aberrations mitogen activated kinase (MAPK) mpk4. further demonstrate, qualitative manner, colocalizations between MPK6 unknown dually phosphorylated MAPK species follow localization associated 65-3 (MAP65-3) telophase microtubular arrays. powerful, versatile adaptable method elucidating spatial relationships compartments. Improved methods sample preparation aiming compensation refractive index mismatches, allow use documentation complex cell elucidation their interactions with proteins.