Ventana Medical Systems, Inc.

Michael J Gizinski

Phillip C Miller

Michael J DeGroff

James A Rybski

Pamela S Vandivort

Anthony L Hartman

The present invention provides an improved method for staining slides using immunochemical reagents. The method comprises the following steps. The assay region of a slide (the region containing the tissue section)is washed with an improved rinsing solution comprising water and a detergent. An evaporation inhibitor liquid is applied to the slide to cover the assay region. For antigens requiring unmasking, the tissue section is combined with an improved, stabilized proteolytic enzyme solution. The slide is rinsed, and the evaporation inhibitor liquid is reapplied to the slide. A primary antibody in an improved diluent containing globulins from the same species as a second antibody is combined with the tissue section for a time sufficient for substantially complete antibody binding. The slide is rinsed, and the evaporation inhibitor liquid is reapplied. A labeled second antibody in the improved diluent is combined with the tissue section for a time sufficient for substantially complete antibody binding. The slide is rinsed, and the evaporation inhibitor liquid is reapplied to the slide. Color development reagents, including a stabilized peroxidase chromophore formulation, are combined with the tissue section for a time sufficient for color development. The stabilized peroxidase chromophore formulation consists essentially of a peroxidase chromophore, at a concentration in the working range of the enzyme, an acidic buffer, a reducing agent, and a glycol. Preferred chromophores include 3,3'-diaminobenzidine and tetrahydrochloride (DAB) and 3-amino-9-ethylcarbazole (AEC). After color development, the tissue section is washed and is then ready for analysis.

Immunohistochemical staining method and reagents therefor

The present invention provides an immunohistochemical staining method using improved reagents. In one aspect, the method uses an evaporation inhibitor liquid to prevent evaporation of reagents applied to an assay region of a slide.

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Tucson, AZ, United States of America

Michael J Gizinski