Immunostaining of DNA in electron microscopy: an amplification and staining procedure for thin sections as alternative to gold labeling. | Zendy

Bernd Bohrmann | Zendy; E. Kellenberger | Zendy

Open Access

Immunostaining of DNA in electron microscopy: an amplification and staining procedure for thin sections as alternative to gold labeling.

Author(s) -

Bernd Bohrmann,

E. Kellenberger

Publication year - 1994

Publication title -

journal of histochemistry and cytochemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.971

H-Index - 124

eISSN - 1551-5044

pISSN - 0022-1554

DOI - 10.1177/42.5.7512586

Subject(s) - uranyl acetate , staining , cryofixation , electron microscope , primary and secondary antibodies , immunogold labelling , microbiology and biotechnology , immunostaining , dna , potassium permanganate , chemistry , biophysics , antibody , biology , biochemistry , immunohistochemistry , optics , genetics , physics , organic chemistry , immunology

We describe a new electron microscopic on-section staining technique with high specificity and sensitivity for DNA-containing structures. Lowicryl HM20 sections of specimens obtained by cryofixation and freeze-substitution are incubated in a first step with a primary IgM antibody specific for double-stranded DNA. The layer of bound antibodies at the section surface is amplified in a successive step by a secondary IgG antibody. Finally, electron scattering of the antibody layer produced is enhanced by staining with a mixture of uranyl acetate and potassium permanganate. The applicability of the method is exemplified by the detection of shape and distribution of various types of bacterial and eukaryotic chromatin.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research