Stereo electron microscope studies of mitotic chromosomes from Chinese hamster ovary cells.

ABSTRACT

Stereo electron microscopy was employed to examine thin sections of Chinese hamster ovary metaphase chromosomes in situ and of chromosomes released from Chinese hamster ovary cells by several methods. Detergent lysis of cells in a buffer containing Mg++ and Ca++ and hypotonic lysis of cells in a hexylene glycol-Ca++ buffer released chromosomes that exhibited a three-dimensional meshwork of about 50 nm chromatin fibers. Fragmentation of cells in serum-free medium by vortexing with glass beads revealed a more dispersed chromosomal morphology with a mesh of 10 to 25-nm fibers exhibiting a presumptive nucleosomal substructure. Possible origins of the various fiber sizes are discussed in terms of current models of metaphase chromosome structure.