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Immunohistochemical staining of bromodeoxyuridine‐labelled cells in the mouse embryo
Author(s) -
Cottell David C.,
Gilmartin Linda,
Bannigan John G.
Publication year - 1989
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1989.tb02937.x
Subject(s) - avidin , staining , biotin , immunohistochemistry , fixation (population genetics) , microbiology and biotechnology , paraformaldehyde , glutaraldehyde , embryo , horseradish peroxidase , chemistry , andrology , biology , pathology , biochemistry , chromatography , immunology , enzyme , medicine , gene
SUMMARY The aim of this study was to compare two immunohistochemical methods, avidin‐biotin peroxidase and immunogold silver staining (IGSS), in the detection of 5‐bromo‐deoxyuridine incorporation in mouse embryo tissues. In addition, two fixation schedules, formal‐saline and a mixture of formaldehyde and glutaraldehyde (4F1G), and two embedding procedures, paraffin wax and the acrylic resin L.R. White, were also compared. Pregnant mice were injected with 600mg per kilogram body weight on days 10 or 15 (plug day = day 1) of gestation and the embryos recovered 2 h after treatment and fixed in formal‐saline or 4F1G. Fixed material was then processed into wax blocks or L. R. White resin. After sectioning, the antigen‐antibody reaction was visualized using either the avidin‐biotin peroxidase or IGSS methods. All methods tested gave a well‐contrasted, highly specific reaction, but IGSS in combination with 4F1G and L. R. White gave a clearer and more sensitive signal than other combinations.