Work

Resinless section electron microscopy of HeLa cell mitotic architecture.

Year: 1986

Type: article

Abstract: The use of resinless sections extends embedment-free electron microscopy to the cytoskeleton of thick specimens. Here we examine HeLa cells rounded at mitosis. Extraction of mitotic HeLa cells with Triton X-100 removes lipids and soluble proteins, leaving the cytoskeletal framework and spindle apparatus. After fixation, the samples are embedded and sectioned, and the temporary embedding resin is removed for direct visualization in the electron microscope. The micrographs show that the cytoskeletal framework, chromosomes, spindle, and centrioles form an interconnected entity. The pericentriolar region, indistinct in conventional micrographs, appears composed of distinct fibers interconnecting the spindle microtubules and centriole. The resinless sections also reveal characteristic lacunae at late anaphase/early telophase. These probably result from reformation of the interphase cytoskeleton lagging reassembly of the nucleus.

Source: Proceedings of the National Academy of Sciences

DOI: 10.1073/pnas.83.23.8996

Authors: B Wagner, G Krochmalnic, S Penman

Institution: Massachusetts Institute of Technology

Language: English

Topic: Microtubule and mitosis dynamics

Subfield: Cell Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funder: National Cancer Institute

Awards: G47087058, G665928895