Staining methods for histology and cytology specimens
First Claim
1. In a method for carrying out the staining of histology or cytology specimens adhered to a slide free of paraffin mounting, which method includes the following steps:
- an optional step of hydrating the specimen adhered to a slide by applying water thereto;
applying excess stain to the specimen;
removing excess stain and/or water by exposing the specimen to a washing-dehydrating reagent;
clearing the specimen of residual washing-dehydrating reagent remaining from a previous washing-dehydrating step and thereby readying the specimen and slide for the application of a cover-slip mountant and a cover-slip;
the improvement comprising;
clearing the specimen by performing a drying-clearing step wherein the specimen is at least partially dried of the washing-dehydrating reagent thereby readying it for the application of a cover-slip mountant and a cover-slip without a chemical-clearing step.
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Abstract
In the field of staining methods for histology and cytology specimens, the improvements of the invention are three-fold. First included is a drying-clearing step whereby a specimen is cleared by drying. Thus, the use of hazardous chemical-clearing agents to remove washing-dehydrating solution prior to cover-slipping is eliminated. Second, in a staining method using spray staining equipment, the use of a low molarity, combination, bluing-washing solution which serves the dual purpose of a washing solution and a bluing reagent, eliminates the use of water in a staining procedure. Third, and also in a staining method using spray staining equipment, the use of a series of sprays of fresh, essentially anhydrous, washing-dehydrating solution to dehydrate a specimen replaces immersion in a series of washing-dehydrating solutions and thereby decreases the amount of chemical consumed and also avoids contamination of the reagent with water and specimen debris.
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Citations
19 Claims
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1. In a method for carrying out the staining of histology or cytology specimens adhered to a slide free of paraffin mounting, which method includes the following steps:
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an optional step of hydrating the specimen adhered to a slide by applying water thereto; applying excess stain to the specimen; removing excess stain and/or water by exposing the specimen to a washing-dehydrating reagent; clearing the specimen of residual washing-dehydrating reagent remaining from a previous washing-dehydrating step and thereby readying the specimen and slide for the application of a cover-slip mountant and a cover-slip; the improvement comprising; clearing the specimen by performing a drying-clearing step wherein the specimen is at least partially dried of the washing-dehydrating reagent thereby readying it for the application of a cover-slip mountant and a cover-slip without a chemical-clearing step. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. In a method for carrying out the staining of histology or cytology specimens whereby the following steps are included:
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exposing a slide having a specimen fixed to it to a washing-dehydrating reagent; removing washing-dehydrating reagent and hydrating the specimen by exposing the slide to an aqueous reagent; staining susceptible elements of the cells of the specimen with hematoxylin by exposing the specimen to an excess of said stain, removing excess stain by exposing the specimen to an aqueous reagent, exposing the specimen to a bluing solution having a pH above 5.0 to turn the hematoxylin blue and readying the specimen for the application of subsequent reagents; removing the excess stain and/or water by exposing the specimen to a washing-dehydrating reagent remaining from a previous washing-dehydrating step and thereby readying the specimen and slide for the application of a cover-slip mountant and a cover-slip; and clearing the specimen of residual washing-dehydrating reagent remaining from a previous washing-dehydrating step and thereby readying the slide for the application of a cover-slip mountant and a cover-slip; the improvement comprising; exposing the slide to a combination bluing-washing solution having a concentration of a basic salt solute of about 0.01 molar yielding a solution with above about pH 5.0, said exposure raising the pH of the specimen above pH 5.0 thereby turning the hematoxylin blue, washing away excess hematoxylin, and readying the specimen for application of a subsequent reagent. - View Dependent Claims (11, 12, 13)
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14. In a method for carrying out the staining of a histology or cytology specimen including the step of applying to a slide having a specimen affixed thereto, a reagent, other than a washing-dehydrating reagent, the improvement comprising:
removing said reagent from the specimen by repeatedly spraying the slide with an essentially anhydrous washing-dehydrating reagent. - View Dependent Claims (15, 16)
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17. A method of readying for the application of a cover-slip mountant and a cover-slip a histology or cytology specimen that has been affixed to a slide and to which stain and/or water has been applied and excess stain and/or water removed by application of a washing-dehydrating reagent, comprising the step of at least partially drying the washing-dehydrating reagent applied to the specimen thereby readying the specimen and slide for application of a cover-slip mountant and cover-slip without a chemical clearing step.
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18. In a method for carrying out the staining of histology or cytology specimens wherein is included the step of staining the susceptible elements of the specimen with hematoxylin wherein an excess of hematoxylin is provided, the improvements comprising:
removing excess hematoxylin by spraying the slide with a combination bluing-washing solution having a concentration of a basic salt solute of no more than about 0.01 molar yielding a solution with above about pH 5.0 and raising the pH of the slide to above pH 5.0 thereby turning the hematoxylin blue and readying the slide for the application of a subsequent reagent. - View Dependent Claims (19)
Specification