Methods and apparatus for the quantitation of nuclear protein
US5202931A · kind A · utility
Assignee
Inventor
Key dates
| Filing date | Jun 23, 1992 |
| Grant date | Apr 13, 1993 |
| Priority date | — |
| Expiry date | Jun 23, 2012 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N1/30
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
An image analysis system is used for the quantitation of nuclear proteins in cell populations. Particularly, the hormonal receptor content of fine needle aspirates of human breast carcinomas are evaluated. Estrogen or progesterone receptors are amplified and visualized in the specimen by a staining technique of the immunoperoxidase type. Monoclonal antibodies specific against the receptor are attached to the receptor sites and are then amplified by a bridging antibody which attaches to the monoclonal antibody and a peroxidase-antiperoxidase complex. A chromagen, diaminobenzidine is combined with the complex and treated with hydrogen peroxide to react with the peroxidase forming an insoluble brown precipitate which marks the receptor sites for optical identification. The specimen is then counterstained with another chromagen, methyl green which is specific to the nucleus of each cell. Two monochromatic filterings optically separate the areas stained by the receptor site optical enhancer and the nuclear area optical enhancer. Measurements of the optical density values of the stained receptor areas yield an intensity value directly related to the quantity of hormonal receptor in the s…
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