Abstract
A simple and versatile technique for the preparation of ultra-thin sections, which can be stained immunohistochemically directly on electron microscope grids, is presented. An anti-hapten immunoperoxidase procedure has been adapted for use on tissue fixed in a purified monomeric glutaraldehyde--picric acid mixture, and embedded in 'L R White', a recently formulated plastic resin. This plastic tolerates the use of partial dehydration of tissue, resulting in higher antigenic yields. In addition, no etching of ultra-thin sections is necessary, and the whole immunostaining procedure can be completed in less than 2 h. A comparison of commonly used fixatives is discussed. High-resolution micrographs showing general staining (uranyl acetate--lead citrate) of rat pancreas, and immunostaining of insulin and TSH in storage granules in perfusion-fixed rat tissue and of lambda-chain immunoreactive cells in immersion-fixed human tonsil are included as examples.
MeSH Terms
Animals
Humans
Immunoglobulin lambda-Chains/metabolism
Immunologic Techniques
Insulin/metabolism
Islets of Langerhans/ultrastructure
Microscopy, Electron/methods
Palatine Tonsil/ultrastructure
Pancreas/ultrastructure
Plastics
Rats
Thyrotropin/metabolism
Chemicals
Immunoglobulin lambda-Chains
Insulin
Plastics
Thyrotropin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Newman G R
Jasani B
Williams E D
References (16)
16 references, click to expand
-
Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures.
J Histochem Cytochem. 1981 Apr;29(4):577-80
PMID: 6166661
-
Simultaneous ultrastructural demonstration of multiple peptides in endocrine cells by a novel immunocytochemical method.
Nature. 1979 Dec 13;282(5740):743-6
PMID: 229417
-
Use of monoclonal antihapten antibodies for immunolocalisation of tissue antigens.
J Clin Pathol. 1981 Sep;34(9):1000-2
PMID: 7024320
-
Enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue.
J Histochem Cytochem. 1981 May;29(5):663-71
PMID: 6166664
-
Immunoperoxidase staining of primate pituitaries with antibodies against the beta subunits of human pituitary glycoprotein hormones.
J Histochem Cytochem. 1981 Jul;29(7):837-43
PMID: 6790604
-
Electron microscopic study of the adrenocorticotropin-producing cell with the use of unlabeled antibody and the soluble peroxidase-antiperoxidase complex.
J Histochem Cytochem. 1972 Aug;20(8):590-603
PMID: 4114593
-
Light and electron microscopic localization of antigens in tissues embedded in polyethylene glycol with a peroxidase labeled antibody method.
J Histochem Cytochem. 1972 Dec;20(12):969-74
PMID: 4640967
-
Immunocytochemical localization of mitochondrial proteins in the rat hepatocyte.
J Histochem Cytochem. 1982 Feb;30(2):139-47
PMID: 7061817
-
Ultrastructural localization of intracellular antigens by the use of protein A-gold complex.
J Histochem Cytochem. 1978 Dec;26(12):1074-81
PMID: 366014
-
Detection of immunoglobulin A by immunofluorescence in glycol methacrylate-embedded human colon.
J Histochem Cytochem. 1982 Jun;30(6):532-5
PMID: 7047638
-
High resolution light and electron microscopic localization of tubulin with the IGS (immuno gold staining) method.
Cell Biol Int Rep. 1981 Sep;5(9):889-99
PMID: 7028278
-
An antiglobulin reagent labelled with colloidal gold for use in electron microscopy.
Immunochemistry. 1974 Aug;11(8):521-2
PMID: 4218593
-
Glutaraldehyde--its purity and stability.
Histochemie. 1972;30(2):162-7
PMID: 5057424
-
Masking of protein antigen by modification of amino groups with carbobenzoxychloride (benzyl chloroformate) and demasking by treatment with nonspecific protease.
J Histochem Cytochem. 1978 Nov;26(11):914-20
PMID: 82572
-
Immunocytochemical localization of galactosyltransferase in HeLa cells: codistribution with thiamine pyrophosphatase in trans-Golgi cisternae.
J Cell Biol. 1982 Apr;93(1):223-9
PMID: 6121819
-
Hydroxyethylmethacrylate embedding: an improved technique.
Med Lab Sci. 1981 Mar;38(2):121-2
PMID: 7040897