Three-dimensional immunogold labeling of nuclear matrix proteins in permeabilized cells

Arjan de Graaf*, Bruno M. Humbel, Nico Stuurman, Paul M.P. van Bergen en Henegouwen, Arie J. Verkleij

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

A preembedment labeling procedure is described for the three-dimensional (3D) labeling of nuclear matrix proteins in permeabilized cells. The procedure is based on the use of ultra-small (1 nm) gold particles as a marker system. This marker penetrates the nucleus more efficiently than the conventionally used 5-10 nm colloidal gold probes. Dehydration is performed by freeze-substitution to preserve the ultrastructure of the cell as optimally as possible. During freeze-substitution the samples are stained by uranyl ions to stain the cellular material throughout the resin section. The 3D gold-labeled and uranyl-stained specimen is embedded in Epon resin and semi-thin (0.2-0.5 m) sections are made for stereo electron microscopy. The applicability of this method is illustrated by the localization of nuclear matrix-associated nuclear bodies in permeabilized interphase and mitotic HeLa cells.

Original languageEnglish
Pages (from-to)827-836
Number of pages10
JournalCell Biology International Reports
Volume16
Issue number8
DOIs
Publication statusPublished - 1992

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