Figure 4

Three-dimensional (3D) reconstruction of immunofluorescence-labelled (lamin AC) isolated cell nuclei in X-, Y- and Z-axes. The images reveal representative cases with type1 and type2 NI. For the type1 intranuclear inclusion 161 optical sections of 0.1 µm were imaged and 113 of them were used for 3D reconstruction. The type2 intranuclear inclusion was reconstructed from 173 of 213 optical Sects. (0.1 µm). The type1 intranuclear inclusion is limited by a lamin AC (nuclear membrane marker) immunopositive membrane. The inclusion is completely closed in all planes and there is no DAPI (blue) staining within it. Lamin AC-positive immunostaining (light green) is also present in the nuclear membrane. The images of the type2 intranuclear inclusion in the bottom row show only immunopositivity for lamin AC in the nuclear membrane while the nucleoplasm appears “empty” in all planes.