[A practical method of whole mount TEM sample preparation and the study of nuclear matrix-intermediate filament scaffolds in K562 cells].
We applied culture plates with bored holes covered with FORMVAR and coated with carbon, to replace the gold grids used in conventional whole mount TEM study. Human erythroleukemia cell line K562 cells plated on the plate surface were extracted using a modified protocol to study scaffolds of nuclear matrix (NM) and intermediate filament (IF). The NM of the K562 cells was found to be composed of interweaving filaments of different diameters, while the cytoplasmic IF were mainly distributed in a radialized pattern. Compared with other techniques of whole mount TEM sample preparations, this one is much more practical and economical, yielding clear NM-IF structures with few artifacts. Study of NM-IF scaffolds in K562 cells might provide a basis for further elucidation of the involvement of NM-IF in the denucleation of mammalian erythroblasts.
Ma
W
W
Institute of Basic Medical Sciences, CAMS and PUMC, Beijing.
Xue
S
S
D006801:Humans; D007382:Intermediate Filaments / Q000648:ultrastructure*; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology*; D008854:Microscopy, Electron; D015530:Nuclear Matrix / Q000648:ultrastructure; D013048:Specimen Handling; D014407:Tumor Cells, Cultured