Histochemical Assay of Œstrogen Receptor

Penney, G. C.; Hawkins, R. A.

doi:10.1016/s0140-6736(80)90858-2

Cited by 5 publications

(2 citation statements)

References 3 publications

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“…Various kinds of fixatives have been used in immunohistochemistry, however great difficulty is encountered when working with highly diffusible antigens such as steroids. Fixatives that cause precipitation of proteins in tissue such as alcohols and acetone have been reported to be ineffective for fixation of such compounds ( 7,8,9 ). Fixatives that act on tissues chemically by forming cross-links between proteins such as aldehydes were reported to be suitable for immunohistochemical staining.…”

Section: Results and Discussion Fixation And Embedding Processesmentioning

confidence: 99%

Immunohistochemical Detection of Steroid Binding in Rabbit Cornea

Rojanasakul

Robinson²

1988

Journal of Ocular Pharmacology and Therapeutics

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An indirect immunoperoxidase technique was used to demonstrate progesterone binding in rabbit cornea as a measure of steroid absorption into corneal tissues. Tissue preparation prior to immune staining was prepared by both a pre-fixed, paraffin-embedding method and a post-fixed, cryostat sectioning method. Rabbit antiprogesterone antiserum and peroxidase conjugated goat antirabbit antiserum were used as a primary and secondary antibody respectively. The immune staining of the antibody-treated sections was then developed in a solution of 3-3' diaminobenzidine-hydrogen peroxide system. The resulting, brown colored, end-products at the site of reaction indicate the location of bound steroid in the sections. Paraffin-embedded sections showed negative staining in both steroid treated and untreated control corneas while fixed frozen sections showed only positive staining for the treated corneas. The results suggest the preservation of the immunogenicity in the treated frozen sections. The negative result observed in the paraffin-embedded sections may be attributed to the loss of steroid from the tissues during the dehydration and paraffin embedding processes.It was found that the immunoreactive progesterone localized mainly in the epithelial cells and the bottom portion of the stroma. Prior application with an excess of triamcinolone acetonide, to the treated corneas gave negative results indicating the reversible and competitive nature of the binding.

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Section: Results and Discussion Fixation And Embedding Processesmentioning

confidence: 99%

Immunohistochemical Detection of Steroid Binding in Rabbit Cornea

Rojanasakul

Robinson²

1988

Journal of Ocular Pharmacology and Therapeutics

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“…Pertschuk et al fixed sections after exposure to oestradiol-fluorescein conjugate. Penney and Hawkins (1980) reported that exposure of rat uterus to acetone for 4 min. resulted in almost total abolition of receptor activity, but have subsequently found this to be incorrect (personal communication).…”

Section: Discussionmentioning

confidence: 99%

The cytochemical demonstration of oestrogen receptors in human breast carcinomas

Walker¹

1981

The Journal of Pathology

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A cytochemical method has been used to demonstrate oestrogen receptor in histological sections of breast carcinomas, and has given similar qualitative results to the biochemical dextra-coated charcoal method. Two conjugation techniques were employed to couple horseradish peroxidase to 17 beta-oestradiol-6-O-carboxymethyloxime-bovine serum albumin. The periodate conjugation method led to a greater degree of coupling but a higher concentration was required for demonstration of oestrogen receptor than that needed when the two-stage glutaraldehyde conjugation method was use, even though this resulted in a lesser degree of coupling. Care in the preparation of tissue has been found to be essential. Formalin-fixed, paraffin-embedded tissue was unsuitable; rapid freezing of fresh tissue, only brief air-drying or acetone fixation of tissue sections and short-term storage were important. A correlation has been shown between the presence of oestrogen receptor in breast carcinomas and good histological differentiation. The poorer differentiated tumours were also found to contain fewer reactive cells.

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Fluorescent cytochemical oestrogen receptor assay: Is it valid in breast cancer?

Furnival

Chong

Watson

et al. 1983

Journal of British Surgery

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The performance of a fluorescent cytochemical oestrogen receptor (ER) assay has been compared with an established dextran-coated charcoal ER assay in 85 cases of primary breast cancer. Both assays were performed on tissue samples from the same tumour, by independent observers in different laboratories. A good qualitative correlation between the two methods was found, with agreement in 92 per cent of cases (P less than 0.0005). Criticisms of the fluorescein-labelled oestrogen methods are discussed. It is concluded that such methods should be validated in each laboratory by comparison with a conventional, quantitative ER assay before being used for clinical decisions.

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Histochemical Assay of Œstrogen Receptor

Cited by 5 publications

References 3 publications

Immunohistochemical Detection of Steroid Binding in Rabbit Cornea

Immunohistochemical Detection of Steroid Binding in Rabbit Cornea

The cytochemical demonstration of oestrogen receptors in human breast carcinomas

Fluorescent cytochemical oestrogen receptor assay: Is it valid in breast cancer?

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