Stabilized two-part hematoxylin solution utilizing pH adjustment
AssigneeVentana Medical Systems, Inc.
InventorsEdward E. Durrant, Christopher A. Koerber
Life of this patent
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Abstract
The present disclosure provides stabilized hematoxylin formulations having a pH of less than 2.4. In general, the stabilized hematoxylin formulations comprise a solvent, a hematoxylin dye, and an acid. The present disclosure also provides methods of using such stabilized hematoxylin formulations to stain biological samples. The present disclosure also provides for methods of readjusting the pH of a stabilized hematoxylin formulation to provide a hematoxylin staining solution.
Claims17
A method of staining a biological sample comprising:
- (a) providing a first reservoir comprising a stabilized hematoxylin formulation having a pH of less than 2.4, wherein the stabilized hematoxylin formulation comprises a hematoxylin dye and an acid in an amount ranging from 0.2% to 4% by total volume of the stabilized hematoxylin formulation;
- (b) mixing an aliquot of the stabilized hematoxylin formulation from the first reservoir with a base to increase the pH of the aliquot of the obtained stabilized hematoxylin formulation to form a hematoxylin staining solution having a pH ranging from 2.4 to 2.6, wherein the base is provided in a second reservoir, and wherein the hematoxylin staining solution is formed in a third reservoir;
- and (c) after forming the hematoxylin staining solution having the pH ranging from 2.4 to 2.6 in the third reservoir, dispensing with an automated staining apparatus the formed hematoxylin staining solution having the pH ranging from 2.4 to 2.6 to the biological sample.
A method of staining a biological sample, comprising:
- (a) dispensing to the biological sample with an automated staining apparatus a stabilized hematoxylin formulation comprising a hematoxylin dye and an acid in an amount ranging from 0.1% to 10% by total volume of the stabilized hematoxylin formulation, wherein the stabilized hematoxylin formulation has a pH of less than 2.4;
- (b) simultaneous or subsequent to the dispensing of the stabilized hematoxylin formulation to the biological sample, dispensing to the biological sample with an automated staining apparatus a readjustment solution, wherein the readjustment solution comprises a strong base;
- (c) permitting the stabilized hematoxylin formulation and the readjustment solution to form a hematoxylin staining solution having a pH ranging from 2.4 to 2.6 while in contact with the biological sample.
A method of staining a biological sample, comprising:
- (a) introducing a stabilized hematoxylin formulation having a pH of less than 2.4 to a container of an automated staining system;
- (b) introducing a readjustment solution to the container of the automated staining system, wherein the readjustment solution comprises a strong base;
- (c) permitting, in the container of the automated staining system, the stabilized hematoxylin formulation and the readjustment solution to form a hematoxylin staining solution having a pH ranging from 2.4 to 2.6;
- and (d) dispensing the formed hematoxylin staining solution from the container of the automated staining apparatus to the biological sample.
Dependencies are inferred from the claim text.