Correlative light and electron microscopic immunocytochemistry on the same section with colloidal gold.

Mar, Henderson; Tsukada, Takashi; Gown, A. M.; Wight, Thomas N.; Baskin, Denis G.

doi:10.1177/35.4.3546488

Cited by 21 publications

(13 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These treatments did not affect the morphological preservation of the tissue and provided efficient and reliable immunostaining at the LM and EM levels. Because of the moderate action of the etching agent, neither semithin nor thin sections needed to be re-embedded according to the protocol of previous studies (Lackie et al 1985;Mar et al 1987;Brorson and Skjörton 1995). Finally, in addition to a direct correlation between immunohistochemistry and immunocytochemistry, we were able to demonstrate immunogold labeling of antigenic sites at the EM level over an exceptionally wide area of tissue perfectly preserved in its morphology.…”

mentioning

confidence: 95%

Re-evaluation of Epoxy Resin Sections for Light and Electron Microscopic Immunostaining

Groos

Reale

Luciano

2001

J Histochem Cytochem.

View full text Add to dashboard Cite

Epoxy resins provide optimal tissue morphology at both the light and the electron microscopic level and therefore enable correlative studies on semithin and thin sections from the same tissue block. Here we report on an approach to retain these advantages for immunolabeling studies by adapting and combining well-known techniques, i.e., surface etching with sodium ethoxide and heat-mediated antigen retrieval. We propose a simple procedure for immunostaining semithin and thin epoxy resin sections. To check its applicability, well characterized, commercially available antibodies (against E-cadherin, alpha-catenin, and beta-catenin) were used on sections of human small intestine. By light microscopy, the immunostaining efficiency was compared on cryo-, paraffin, and epoxy semithin sections processed in parallel. The most detailed results were obtained on semithin sections, where the labeling precisely delineated the lateral plasma membrane of the enterocytes. At the electron microscopic level the procedure did not damage the structures and allowed an efficient, reproducible immunogold labeling extending homogeneously over exceptionally wide tissue areas. The three antibodies specifically labeled the zonula adherens of the junctional complex between epithelial cells and, in agreement with light microscopic observations, the lateral plasma membrane.

show abstract

mentioning

confidence: 95%

Re-evaluation of Epoxy Resin Sections for Light and Electron Microscopic Immunostaining

Groos

Reale

Luciano

2001

J Histochem Cytochem.

View full text Add to dashboard Cite

show abstract

“…These three methods each have advantages and disadvantages, and researchers are required to select the most effective method appropriate to the tissues under observation. Pre-embedding immunocytochemical staining has provided successful observations by light microscopy and electron microscopy in the same specimen [9, 16]. However, permeabilization treatments are sometimes necessary to obtain optimal labeling of intracellular antigenic sites, particularly with thick sections, such as vibratome sections [7, 9, 14].…”

Section: Discussionmentioning

confidence: 99%

“…Although Mar et al . [9] described immunocytochemical observation of rat growth hormone-containing cells between light microscopic images and electron microscopic images of the same cell, they used one plastic-embedded sample, but not a paraffin-embedded sample.…”

mentioning

confidence: 99%

Immunoelectron Microscopic Observation of Chicken Glucagon-Like Peptide (GLP)-1-Containing Cells in Tissues Derived from Thin Section, Paraffin Block and Conventional Method

Watanabe

Nishimura

Monir

et al. 2014

The Journal of Veterinary Medical Science

View full text Add to dashboard Cite

The purpose of the present study was to investigate the possibility of immunoelectron microscopic observation of endocrine cells in paraffin-embedded tissues. The procedure, which involves reprocessing from sliced tissues and immunohistochemical staining by colloidal-gold immunolabeling of paraffin sections from paraffin blocks, was able to reveal the fine characteristics of secretory granules containing glucagon-like peptide-1. Morphometric analyses of the secretory granules showed no significant differences between the reprocessing procedure and a conventional post-embedding procedure, which was performed as a control. The reprocessing procedure has some advantages besides providing information on secretory granules containing the amino acid peptide. For example, the same cell can be observed under both a light microscope and the electron microscope. In addition, the high-electron densities of silver-enhanced gold particles are easily recognized, and the boundary between the profile of the granules and the immunogold labeling is clearly shown at the electron microscopic level. Furthermore, the procedure, which is inexpensive and does not require special devices, can effectively use precious samples that are already paraffin-embedded and unable to be obtained twice, such as the case for endangered animals and rare pathological tissues. To the best of our knowledge, the present study is the first to report the advantages of the reprocessing method for sliced paraffin sections of gut endocrine cells.

show abstract

“…Briefly, before immunostaining slides with 1-2-pm plastic sections were submerged in a solution of saturated sodium hydroxide in absolute alcohol (sodium ethoxide), diluted 1:1 with fresh absolute alcohol, for 15-20 min at room temperature, and were processed for immunocytochemical staining as previously described (Snow et al, 1988;Mar et al, 1987).…”

Section: Methodsmentioning

confidence: 99%

Peripheral distribution of dermatan sulfate proteoglycans (decorin) in amyloid-containing plaques and their presence in neurofibrillary tangles of Alzheimer's disease.

Snow¹,

Mar²,

Nochlin³

et al. 1992

J Histochem Cytochem.

Self Cite

141

View full text Add to dashboard Cite

We used a polydonal antibody and a mixture of three monoclonal antibodies (MAb), all recognizing the protein core of the s m a l l dermatan sulfate proteoglycan (DSPG) (known as PG-II or decorin) derived from human skin fibroblasts, to immunolocalize this molecule in the characteristic lesions in Alzheimer's brain. All antibodies demonstrated positive decorin immunoStaining in both the amyloid deposits of neuritic plaques (NPs) and the filamentous structures within neurofbrillaty tangles (NFTs). Unlike heparan sulfate proteoglycans (HSPGs), which tend to be evenly distributed throughout NPs containing amyloid fibrils, decorin was pri-

show abstract

Correlative light and electron microscopic immunocytochemistry on the same section with colloidal gold.

Cited by 21 publications

References 22 publications

Re-evaluation of Epoxy Resin Sections for Light and Electron Microscopic Immunostaining

Re-evaluation of Epoxy Resin Sections for Light and Electron Microscopic Immunostaining

Immunoelectron Microscopic Observation of Chicken Glucagon-Like Peptide (GLP)-1-Containing Cells in Tissues Derived from Thin Section, Paraffin Block and Conventional Method

Peripheral distribution of dermatan sulfate proteoglycans (decorin) in amyloid-containing plaques and their presence in neurofibrillary tangles of Alzheimer's disease.

Contact Info

Product

Resources

About