Post-embedding immunoperoxidase staining of glial fibrillary acidic protein for light and electron microscopy

DeArmond, Stephen J.; Siegel, Matthew; Dixon, Richard F.; Eng, Lawrence F.

doi:10.1016/0165-5728(81)90003-5

Cited by 20 publications

(7 citation statements)

References 17 publications

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“…Abundant bundles of 10 nm filaments are the most characteristic components of Q 1993 Wiley-Liss, Inc. fibrous astrocytes, whereas these are much less numerous in protoplasmic astrocytes (Peters et al, 1991). Therefore, with anti-GFAP, protoplasmic astrocytes stain not at all or stain less intensely than fibrous astrocytes (Dearmond et al, 1981).…”

Section: Introductionmentioning

confidence: 99%

Distribution of metallothionein in the human central nervous system

Blaauwgeers¹,

Smitt

Jong

et al. 1993

Glia

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The distribution of metallothionein (MT), a metal-binding protein, was examined immunohistochemically in the normal human brain and spinal cord. Paraffin-embedded brain tissue from three patients who had died from a non-neurological disease and were free of histopathological central nervous system alterations were processed. The results of the present study demonstrate that MT is readily detectable in a subgroup of astrocytes in the normal human brain. MT staining is most intense on grey matter astrocytes that bear short stout processes and which probably represent protoplasmic astrocytes. Using anti-MT and anti-glial fibrillary acidic protein immunostaining, we could demonstrate two subpopulations of astrocytes that were mutually exclusive. The functional significance of MT-expression in protoplasmic astrocytes is not entirely clear. Metal detoxification is only one of the many postulated functions of MT. The finding that staining for MT permits subtyping of astrocytes may be of great importance in glia research and surgical pathology of the human brain.

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Section: Introductionmentioning

confidence: 99%

Distribution of metallothionein in the human central nervous system

Blaauwgeers¹,

Smitt

Jong

et al. 1993

Glia

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“…De Armond et al (1981) have shown that NaOH or H202 dissolves Araldite with an action which resembles a deparaffination. Their technique involves the preparation of NaOH saturated ethanol which is then diluted in a proportion 1:3 -NaOH saturated ethanol (1) ethanol (3); this mixture is then employed to dissolve the Araldite.…”

Section: Discussionmentioning

confidence: 98%

“…Firstly, they dissolve, as in the case of Araldite (de Armond et al, 1981), the embedding medium, and secondly, by permeabilization of the tissues they permit the penetration of the antibodies. Antibody penetration is obvious throughout the semithin sections whereas only 3-5 ~m penetration was obtained with sections cut on a vibratome or cryostat.…”

Section: Discussionmentioning

confidence: 99%

“…As with other embedding techniques, i.e. Araldite (de Armond et al, 1981;Berod et al, 1984), a mixture of glycol methacrylate and Queto1523 (Takamiya et al, 1979) or Spur (Seitz et al, 1986), a pretreatment of the sections was necessary to obtain immunolabelling. In the case of Quetol 651 this pre-treatment is carried out with ethanol containing a hydroxide.…”

Section: Discussionmentioning

confidence: 99%

“…A minimum thickness of 200 nm is required in order to observe immunolabelled sites revealed by immunoperoxidase (de Armond et al, 1981) with ease.…”

Section: Discussionmentioning

confidence: 99%

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The use of Quetol 651 for the post-embedding immunohistochemical demonstration ofγ-aminobutyric acid on semithin sections

Denizot

Arnold²,

Geffard

et al. 1988

Histochem J

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Quetol 651 was used as an embedding medium for the demonstration of gamma aminobutyric acid (GABA) in semithin sections by the peroxidase-anti-peroxidase method. In order to demonstrate the immunoreactivity, the embedding medium was partially dissolved using absolute ethanol containing 0.8-1 M NaOH or KOH for 5-7 min. The experimental procedure was elaborated by testing the GABAergic sites in the endings surrounding the small neurones of the anterior exterolateral nucleus of a mormyrid fish and in the pyramidal cells of the electrosensory lateral line lobe of gymnotoid fish by applying anti-GAD (glutamic acid decarboxylase) antiserum. To test the general validity of the use of Quetol 651, GABAergic sites were also identified in the central nervous system of an insect, the honey bee, with anti-GABA and anti-GAD antisera. The intensity of labelling revealed by immunoperoxidase applied to Quetol 651-embedded semithin sections, demonstrated high precision and gave good resolution for light microscopical observations.

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