Immunohistochemical staining method and reagents therefor
AssigneeVentana Medical Systems, Inc.
InventorsPhillip C. Miller, Michael J. Degroff, Michael J. Gizinski, James A. Rybski, Pamela S. Vandivort
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Abstract
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.
Claims12
A method of preventing evaporation of reagents applied to an assay region of a slide, said assay region comprising an aqueous phase reagent, said method comprising covering said assay region with an evaporation inhibitor liquid said evaporation inhibitor liquid comprising a liquid which is immiscible with said aqueous phase reagent, has a density less than that of said aqueous phase reagent, and does not interfere with reactions taking place in said aqueous phase reagent.
An improved assay method wherein an assay reagent or sample is bound to an assay region of a slide, the improvement comprising covering said assay region with an evaporation inhibitor liquid consisting essentially of an oil of the medium chain alkane family.
An improved immunohistochemical method comprising, in the following order:
- (a) rinsing a tissue section with an aqueous rinsing solution consisting essentially of a substantially salt-free water and a sufficient amount of a nonionic biological detergent to reduce surface tension to provide for even sheeting of the solution;
- (b) applying a sufficient amount of an evaporation inhibitor liquid to cover said tissue section;
- (c) combining a primary antibody in a diluent consisting essentially of a physiologic buffer and the globulin fraction of nonimmune serum from the same species as a second antibody at a protein concentration sufficient to inhibit nonspecific antibody binding with said tissue section for a time sufficient for substantially complete antibody binding;
- (d) rinsing said tissue section with a rinsing solution consisting essentially of a substantially salt-free water and a sufficient amount of a nonionic biological detergent to reduce surface tension to provide for even sheeting of the solution;
- (e) applying a sufficient amount of an evaporation inhibitor liquid to cover said tissue section;
- (f) combining a labeled second antibody in a diluent consisting essentially of a physiologic buffer and the globulin fraction of nonimmune serum from the same species as said second antibody at a protein concentration sufficient to inhibit nonspecific antibody binding with said tissue section for a time sufficient for substantially complete antibody binding;
- (g) rinsing said tissue section with a rinsing solution consisting essentially of a substantially salt-free water and a sufficient amount of a nonionic biological detergent to reduce surface tension to provide for even sheeting of the solution;
- (h) applying a sufficient amount of an evaporation inhibitor liquid to cover said tissue section;
- (i) combining color development reagents with said tissue section for a time sufficient for color development;
- and
- (j) rinsing said tissue section with a rinsing solution consisting essentially of a substantially salt-free water and a sufficient amount of a nonionic biological detergent to reduce surface tension to provide for even sheeting of the solution.
Dependencies are inferred from the claim text.