Immunoelectron microscopic demonstration of thyroglobulin and thyroid hormones in rat thyroid gland.

Ring, P.; Johanson, Viktor

doi:10.1177/35.10.2442241

Cited by 11 publications

(1 citation statement)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…overnight. Both, heat and catalyst curing may have deleterious effects on the structure of nervous but also of other tissues, such as human prostate [25] and breast cancer [26], rat thyroid gland [27], pancreas [28 , 29], and testes [30]. Poor tissue preservation in these instances may have occurred possibly because the resin polymerized too rapidly, which can cause considerable external contraction of the blocks and disrup tive tissue swelling inside [20,29].…”

Section: Discussionmentioning

confidence: 99%

Use of LR White Resin for Post-Embedding Immunolabelling of Brain Tissue

Gocht¹

1992

Cells Tissues Organs

View full text Add to dashboard Cite

The object of this study was to investigate the applicability of the acrylic resin ‘LR White’ to immunolabelling of various antigenic determinants in aldehyde-fixed rat CNS tissue. Antibodies were used, which worked well in paraffin sections and therefore were suitable to detect antigens resistant to complete dehydration and heat. Different LR White embedding protocols were employed in order to select the preparation conditions that adequately preserved both the antigenicity and fine structure. Specimens were completely dehydrated with up to 100% ethanol, which was followed by various infiltration times with LR White monomer. Polymerization of the resin was induced by heat, a chemical catalytic procedure (accelerator), or ultraviolet (UV) light. Paraffin, as well as semithin and ultrathin LR White sections were incubated with antibodies reacting to antigens located on the cell surface (stage-specific embryonic antigen-1; SSEA-1), within the plasma membrane (myelin basic protein), in the cytosol (HNK-1, S100 protein), in the cytoskeleton (GFAP, vimentin, neurofilament protein, INT-FIL), and in the extracellular matrix (laminin). All of the examined antigens were immunocytochemically detectable in paraffin-embedded material, while the carbohydrate moieties, HNK-1 and SSEA-1, were not immunoreactive in LR White sections. However, in cryostat sections processed for pre-embedding immunoelectron microscopy, the HNK-1 epitope and SSEA-1 were immunolabelled. Polymerization carried out under UV light led to better structural preservation of brain tissue than resin cured with heat or catalyst. The length of prior infiltration with monomer apparently had no effect on tissue preservation. Consequently, UV light-induced polymerization of LR White gives acceptable morphology of brain tissue. However, the use of this acrylic resin is restricted to the detection of some CNS antigens only.

show abstract

Section: Discussionmentioning

confidence: 99%

Use of LR White Resin for Post-Embedding Immunolabelling of Brain Tissue

Gocht¹

1992

Cells Tissues Organs

View full text Add to dashboard Cite

show abstract

Embedding Media for Section Immunocytochemistry

Griffiths

1993

Fine Structure Immunocytochemistry

View full text Add to dashboard Cite

Localization of GABA and glycine in goldfish retina by electron microscopic postembedding immunocytochemistry: improved visualization of synaptic structures with LR White resin

Studholme

Yazulla

1988

J Neurocytol

View full text Add to dashboard Cite

A post-embedding, electron microscopic immunocytochemistry technique, modified from existing protocols, was used to examine the labelling patterns of GABA immunoreactivity and glycine immunoreactivity in goldfish retina. Retinae were fixed in mixed aldehyde solution, dehydrated in ethanol, stained en bloc with uranyl acetate and phosphotungstic acid and embedded in LR White resin. Substances were localized in thin sections by floating grids first on a drop of primary antiserum and then on a colloidal gold-IgG conjugate. Finally, grids were exposed to osmium vapour. The localization of GABA immunoreactivity matched that of [3H]-GABA uptake or glutamate decarboxylase immunoreactivity as described previously. In the outer retina, GABA immunoreactivity was found in the cell bodies and axon terminals of H1 horizontal cells and their dendrites opposite cone photoreceptor terminals. Selected amacrine cell bodies were labelled, as were many processes, both synaptic and non-synaptic, throughout the inner plexiform layer, including most amacrine cell processes contacting the synaptic terminals of type Mb bipolar cells. Numerous amacrine cells, their processes in the inner and outer plexiform layers, and photoreceptor terminals contained glycine immunoreactivity in a distribution similar to that shown by [3H]-glycine uptake. Despite the absence of osmium in the primary or secondary fixative, our protocol results in excellent visibility of synaptic structures and detectability of the colloidal gold immunolabel. Also, it does not cause extraction of the HRP/DAB reaction product and is therefore suitable for double-label analysis of neurons labelled with horseradish peroxidase.

show abstract

Immunoelectron microscopic demonstration of thyroglobulin and thyroid hormones in rat thyroid gland.

Cited by 11 publications

References 9 publications

Use of LR White Resin for Post-Embedding Immunolabelling of Brain Tissue

Use of LR White Resin for Post-Embedding Immunolabelling of Brain Tissue

Embedding Media for Section Immunocytochemistry

Localization of GABA and glycine in goldfish retina by electron microscopic postembedding immunocytochemistry: improved visualization of synaptic structures with LR White resin

Contact Info

Product

Resources

About