Abstract: Improved biotin blocking methods are provided which utilize a high-affinity monomeric biotin-binding protein of the avidin family that have only a single biotin binding site. Because the biotin-binding protein is monovalent, it can be applied as a single reagent. Since it does not introduce any additional biotin binding sites, as would be the case with native tetrameric streptavidin, no further blocking of biotin binding sites is necessary. Furthermore, because it is a single reagent, it can be combined with other steps such as the peroxidase blocking step, and both endogenous peroxidase and endogenous biotin can be blocked simultaneously. The biotin blocking methods may be used in the performance of immunoassays, such as tissue sample containing endogenous biotin is stained by IHC to detect a specific analyte. Prior to staining, the tissue is first blocked for endogenous biotin, according to the present invention, by the application of a high-affinity biotin-binding protein.

Abstract: A sample protection method is provided which may be used for protecting a biological sample on a microscope slide, such as during heat-induced target retrieval and/or after heat-induced target retrieval such that: 1) the sample remains adherent to the microscope slide; and 2) the microscopic morphology of the biological sample remains intact. In some embodiments, the sample protection method may include the steps of: creating a sectioned sample that is in contact with a microscope slide; applying a protecting reagent onto a sectioned sample that is in contact with a microscope slide and drying the protecting reagent in which the protecting reagent may be both applied and dried onto the sectioned sample before and/or after performing target retrieval on the sectioned sample. The protecting reagent may include a water-soluble polymer and/or a water-soluble wax, such as polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, alginic acid, and carrageenan.

Abstract: A method to produce a rapid immunohistochemical detection of an antigen from a biological sample is provided. Preferably, the method may be used for rapid immunohistochemical staining of a biological sample that allows completion of the staining process in less than ten minutes, such as during a cancer removal procedure. In some embodiments, the method may include the steps of: depositing a section of the biological sample on a slide; permeabilizing the section of the biological sample; incubating the section of the biological sample with a secondary antibody having a detectable label; removing unbound secondary antibody from the section of the biological sample; mounting the section of the biological sample; and detecting secondary antibody bound to the section of the biological sample.

Abstract: An antigen protection method for enabling delayed sample processing may include: fixing a sample; embedding the sample; sectioning the sample and placing a section of the sample in contact with a microscope slide to produce a sectioned sample; de-embedding the sectioned sample; optionally rehydrating the sectioned sample; performing target retrieval on the sectioned sample; and optionally performing target visualization on the sectioned sample. The method may further include the steps of: applying a protecting reagent onto the sectioned sample; drying the protecting reagent; and waiting a period of time, such as greater than six hours, and these steps may be performed after the de-embedding step, rehydrating step, and/or target retrieval step.

Abstract: A sample protection method is provided which may be used for protecting a biological sample on a microscope slide, such as during heat-induced target retrieval and/or after heat-induced target retrieval such that: 1) the sample remains adherent to the microscope slide; and 2) the microscopic morphology of the biological sample remains intact. In some embodiments, the sample protection method may include the steps of: creating a sectioned sample that is in contact with a microscope slide; applying a protecting reagent onto a sectioned sample that is in contact with a microscope slide and drying the protecting reagent in which the protecting reagent may be both applied and dried onto the sectioned sample before and/or after performing target retrieval on the sectioned sample. The protecting reagent may include a water-soluble polymer and/or a water-soluble wax, such as polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, alginic acid, and carrageenan.

Abstract: A deparaffinizing composition and method are provided for removing paraffin wax from biological samples. The deparaffinizing composition may include: a paraffin solvent that may be a first apolar paraffin solvent; a second apolar solvent that may be at least partially miscible with water; a first polar solvent that may be miscible with both water and apolar solvents; a second polar solvent that may be water; and a detergent. The deparaffinizing method may include the steps of: incubating the sample for a time period in a deparaffinizing composition; and rinsing the sample with an aqueous buffer. Preferably, the step of incubating the sample for a time period in the deparaffinizing composition may include heating the sample and the composition to between approximately 35 to 65 degrees Celsius.

Abstract: The present disclosure relates to counterstains for staining a biological sample on a single slide in preparation for microscopic examination. The counterstains are used to analyze the sample on the single slide using both brightfield and fluorescent illumination. The counterstains can identify both morphological details and molecular structures within the cells contained in the sample. The counterstains can be used in conjunction with other molecular stains.

Abstract: The present disclosure relates to an aqueous sealing fluid for sealing a stained biological specimen present on a microscope slide for microscopic analysis. The sealing fluid protects the biological specimen from drying artifacts, protects stains performed on biological specimen from fading, and protects stains performed on biological specimens from being dissolved by organic mounting media. Methods of using the aqueous sealing fluid are also disclosed.

Abstract: A method for labeling concentration density differentials of an analyte in a biological sample is provided. The method may including the steps of: binding an enzyme to an analyte contained in a sample, the enzyme capable of acting on at least two chromogens; incubating the sample with a first chromogen for a first time period to generate a first color chromogen-enzyme product, the first color chromogen-enzyme product reflecting light observable as a first color; and incubating the sample with a second chromogen for a second time period to generate a second color chromogen-enzyme product, the second color chromogen-enzyme product reflecting light observable as second color. A combination of the light observable as the first color and the light observable as the second color may be observable as a third color, and each color may describe a different analyte density in the biological sample.

Abstract: The present disclosure relates methods of chromogen layering, wherein a first chromogen and a first stain (color) are produced on a sample, specific for a first analyte, followed by a second chromogen and a second stain (color) being produced on the same sample, specific for a second analyte. In addition, if desired, by overlaying the second stain on top of the first stain, a unique third color is produced that is specific for a third analyte. Therefore, the distribution of different colors throughout the sample could be used to identify at least two or more analytes simultaneously within a single sample.

Abstract: Disclosed are methods for treating a biological sample for an analyte-staining assay. These methods are conducted with the use of a single aqueous multi-functional composition for the purposes of performing the steps of deparaffinization, rehydration, antigen retrieval and endogenous enzyme blocking simultaneously optionally at an elevated temperature. These methods are alternatively conducted with use of a single aqueous multi-functional composition that contains a thermostable protease for the purpose of deparaffinization, rehydration, antigen retrieval and endogenous enzyme blocking at an elevated temperature.

Abstract: A deparaffinizing composition and method are provided for removing paraffin wax from biological samples. The deparaffinizing composition may include: a paraffin solvent that may be a first apolar paraffin solvent; a second apolar solvent that may be at least partially miscible with water; a first polar solvent that may be miscible with both water and apolar solvents; a second polar solvent that may be water; and a detergent. The deparaffinizing method may include the steps of: incubating the sample for a time period in a deparaffinizing composition; and rinsing the sample with an aqueous buffer. Preferably, the step of incubating the sample for a time period in the deparaffinizing composition may include heating the sample and the composition to between approximately 35 to 65 degrees Celsius.

Abstract: A sample processing system that may be automated and methods are disclosed where samples are arranged on a carrier element and a process operation control system automatically processes the samples perhaps robotically with an operationally-influential exteriorly-consequential information monitor or a data capture element. Significant process details as well as operationally-influential exteriorly-consequential information may be monitored and an automatic notice element may cause notification of a person at some display that may be remote. Various people may be notified, such as an administrator, a supplier, or a manufacturer of an opportunity for some action such as reagent reordering or the like. A simulated motion display may be included to “watch” simulated operation in real time or long after completion of the actual processing.

Abstract: A method for labeling concentration density differentials of an analyte in a biological sample is provided. The method may including the steps of: binding an enzyme to an analyte contained in a sample, the enzyme capable of acting on at least two chromogens; incubating the sample with a first chromogen for a first time period to generate a first color chromogen-enzyme product, the first color chromogen-enzyme product reflecting light observable as a first color; and incubating the sample with a second chromogen for a second time period to generate a second color chromogen-enzyme product, the second color chromogen-enzyme product reflecting light observable as second color. A combination of the light observable as the first color and the light observable as the second color may be observable as a third color, and each color may describe a different analyte density in the biological sample.

Abstract: The present disclosure relates methods of chromogen layering, wherein a first chromogen and a first stain (color) are produced on a sample, specific for a first analyte, followed by a second chromogen and a second stain (color) being produced on the same sample, specific for a second analyte. In addition, if desired, by overlaying the second stain on top of the first stain, a unique third color is produced that is specific for a third analyte. Therefore, the distribution of different colors throughout the sample could be used to identify at least two or more analytes simultaneously within a single sample.

Abstract: The present disclosure relates to counterstains for staining a biological sample on a single slide in preparation for microscopic examination. The counterstains are used to analyze the sample on the single slide using both brightfield and fluorescent illumination. The counterstains can identify both morphological details and molecular structures within the cells contained in the sample. The counterstains can be used in conjunction with other molecular stains.

Abstract: A sample processing system 101 that may be automated and methods are disclosed where a number of sample processing systems 101, such as stainer, may be connected to a number of separate full function computers 181 through a stainer network 183 that may be isolated from other communication traffic. A network configuration may permit scalability and addressability so that additional sample processing systems 101, additional separate full function computers 181, and additional other devices such as label printers 200 may be easily added to the system. One or more remote information links 171 may be provided so that information transfer on a continuous or perhaps constant basis can be accommodated.

Abstract: The present disclosure relates to an aqueous sealing fluid for sealing a stained biological specimen present on a microscope slide for microscopic analysis. The sealing fluid protects the biological specimen from drying artifacts, protects stains performed on biological specimen from fading, and protects stains performed on biological specimens from being dissolved by organic mounting media. Methods of using the aqueous sealing fluid are also disclosed.

Abstract: Disclosed are methods for treating a biological sample for an analyte-staining assay. These methods are conducted with the use of a single aqueous multi-functional composition for the purposes of performing the steps of deparaffinization, rehydration, antigen retrieval and endogenous enzyme blocking simultaneously optionally at an elevated temperature. These methods are alternatively conducted with use of a single aqueous multi-functional composition that contains a thermostable protease for the purpose of deparaffinization, rehydration, antigen retrieval and endogenous enzyme blocking at an elevated temperature.

Abstract: The present invention concerns an apparatus for staining tissue samples, said apparatus including a reagent section or reagent containers; at least one staining section or tissue samples, a robotic head or robotic element that may move reagent to a predetermined tissue sample, said robotic element being moveable above the reagent and the staining sections, a control element that may manage a staining process, a 2-D optical sensor to detect two-dimensional image data of a relevant property and that can feed the captured image data to the control element. By providing the robotic element with a 2-D optical sensor, a common image processor may be provided having multiple functions. By using a 2-D optical image processing system, the control system of the apparatus may easily be adapted to read various types of data presentations, just as actual images for sections of the apparatus may be identified in order to assess the condition of the apparatus.