508 results about "Immunohistochemistry" patented technology

Conjugate compositions are disclosed that include a specific-binding moiety covalently coupled to a nanoparticle through a heterobifunctional polyalkyleneglycol linker. In one embodiment, a conjugates is provided that includes a specific-binding moiety and a fluorescent nanoparticle coupled by a heterobifunctional PEG linker. Fluorescent conjugates according to the disclosure can provide exceptionally intense and stable signals for immunohistochemical and in situ hybridization assays on tissue sections and cytology samples, and enable multiplexing of such assays.

A highly sensitive assay is disclosed which combines immunomagnetic enrichment with multiparameter flow cytometric and immunocytochemical analysis to detect, enumerate and characterize carcinoma cells in the blood. The assay can detect one epithelial cell or less in 1 ml of blood and has a greater sensitivity than conventional PCR or immunohistochemistry by 1-2 orders of magnitude. In addition, the assay facilitates the biological characterization and staging of carcinoma cells.

Disclosed are methods and systems for analyzing images of specimens processed by a programmable quantitative assay or more specifically a robust programmable quantitative dot assay, PDQA, that enable specimens to be imaged and assessed across a wide variety of conditions and applications. Specific embodiments directed to immunohistochemical applications provide more quantitative methods of imaging and assessing biological samples including tissue samples.

Apparatus, methods, and computer-readable media are provided for segmentation, processing (e.g., preprocessing and / or postprocessing), and / or feature extraction from tissue images such as, for example, images of nuclei and / or cytoplasm. Tissue images processed by various embodiments described herein may be generated by Hematoxylin and Eosin (H&E) staining, immunofluorescence (IF) detection, immunohistochemistry (IHC), similar and / or related staining processes, and / or other processes. Predictive features described herein may be provided for use in, for example, one or more predictive models for treating, diagnosing, and / or predicting the occurrence (e.g., recurrence) of one or more medical conditions such as, for example, cancer or other types of disease.

Methods for identifying Quantifiable Internal Reference Standards (QIRS) for immunohistochemistry (IHC). Also disclosed are methods for using QIRS to quantify test antigens in IHC.

This invention relates to methods of analyzing a tissue sample from a subject. In particular, the invention combines morphological staining and / or immunohistochemistry (IHC) with fluorescence in situ hybridization (FISH) within the same section of a tissue sample. The analysis can be automated or manual. The invention also relates to kits for use in the above methods.

Scanning and analysis of cytology and histology samples uses a flatbed scanner to capture images of the structures of interest such as tumor cells in a manner that results in sufficient image resolution to allow for the analysis of such common pathology staining techniques as ICC (immunocytochemistry), IHC (immunohistochemistry) or in situ hybridization. Very large volumes of such material are scanned in order to identify cells or clusters of cells which are positive or warrant more detailed examination, and if analysis at higher resolution is necessary, information regarding these positive events is transferred to a secondary microscope, such as a conventional scanning microscope, to allow further analysis and review of the selected regions of the slide containing the sample.

The invention is composed of monoclonal human MN antibodies or MN antibody fragments that target the GEEDLP (SEQ ID NO: 118) repeat within the proteoglycan domain. The proteoglycan domain of the MN cell surface protein contains four of these identical GEEDLP (SEQ ID NO: 118) repeats. Binding to the desired epitope is verified by competition ELISA, where ELISA signal can be attenuated by co-incubation with a peptide containing this repeat (PGEEDLPGEEDLP (SEQ ID NO: 119)). This inhibition of binding can also be verified using Biacore assays, where binding of desired antibodies to immobilized MN or proteoglycan peptides can be inhibited by the peptide repeat. In addition to binding to the peptide repeat, human anti-MN antibodies can inhibit the cell adhesion of CGL-1 cells to MN coated plastic plates. Human anti-MN antibodies have been used to diagnose and quantify MN expression in cancer cells and tumors using FACS and immunohistochemical methods. An example is also provided where a human anti-MN IgG1 mediates tumor cell lysis though antibody-dependent cell-mediated cytotoxicity. Therefore, these antibodies will be useful for the treatment of cancers in which MN is upregulated or can be useful for the diagnosis of cancers in which MN is upregulated.

The present disclosure describes an IHC assay for detecting and quantifying spatially proximal pairs of PD-1-expressing cells (PD-1+ cells) and PD-Ligand-expressing cells (PD-L+ cells) in tumor tissue, and the use of the assay to generate proximity biomarkers that are predictive of which cancer patients are most likely to benefit from treatment with a PD-1 antagonist. The disclosure also provides methods for testing tumor samples for the proximity biomarkers, as well as methods for treating subjects with a PD-1 antagonist based on the test results.

Antibody / signal-generating moiety conjugates are disclosed that include an antibody covalently linked to a signal-generating moiety through a heterobifunctional polyalkyleneglycol linker. The disclosed conjugates show exceptional signal-generation in immunohistochemical and in situ hybridization assays on tissue sections and cytology samples. In one embodiment, enzyme-metallographic detection of nucleic acid sequences with hapten-labeled probes can be accomplished using the disclosed conjugates as a primary antibody without amplification.

A method of in situ immunohistochemical analysis of a biological sample is provided. The method allows for the multiplex and simultaneous detection of multiple antigens, including multiple nuclear antigens, in a tissue sample.

Automated sample processing systems may include onboard efficient high-speed mixing of at least two components with an automatic vertical force fluidic turbulent component mixer of which a mixed component may be aspirated and high-speed dispensed in a mixing vial. Other aspects may include single sweep applying a multi-treatment cleaning cycle to at least one slide. A multi-treatment cleaning cycle may include a washing treatment and a drying treatment. In yet other aspects the present invention may include an automated recovery sample processing system with the capability of detecting at least one immediate condition of a fortuitously terminated automatic sample processing run and perhaps even an automatic terminated sample processing run reconstruction calculator.

The present invention provides monoclonal antibodies (MAbs) selective for the N-termini and C-termini of the gastrin hormone forms, gastrin-17 (G17), glycine-extended gastrin-17 (G17-Gly), gastrin-34 (G34) and glycine-extended gastrin-34 (G34-Gly); and the hybridomas that produce these MAbs. Also provided are panels of MAbs useful for the detection and quantitation of gastrin-17 (G17), glycine-extended gastrin-17 (G17-Gly), gastrin-34 (G34) and glycine-extended gastrin-34 (G34-Gly). These assays are useful for monitoring a gastrin-mediated disease or condition, or for monitoring the progress of a course of therapy. The invention further provides solid phase assays including immunohistochemical (IHC) and immunofluorescence (IF) assays suitable for detection and visualization of gastrin species in solid samples, such as biopsy samples or tissue slices. Pharmaceutical compositions of the MAbs of the invention are also provided, along with methods of diagnosis, prevention and treatment of gastrin-mediated diseases or conditions. Methods of evaluating a gastrin hormone-blocking treatment are described. The course of a gastrin-mediated disease or condition may be monitored in a patient by means of assay methods provided.

A computerized method for immunohistochemistry analysis of tissue utilizes digital images of multiple adjacent tissue sections aligned within a computerized software and processed with an algorithm to quantify a two-dimensional IHC signature score for each respective slide image. In various embodiments, the IHC score is performed over several adjacent sections and further processed to produce a three-dimensional IHC quantification referred to as an IHC signature map.

Provided is an IL-18 receptor antibody usable for immunohistochemistry, which functions as an IL-18 receptor antagonist. Also provided is a pharmaceutical composition and method for preventing and / or treating an IL-18-dependent disorder, particularly skin thickening caused by ultraviolet, using the antibody. An antibody against the IL-18 receptor α subunit, which is characterized by binding specifically to a polypeptide consisting of the amino acid sequence shown by SEQ ID NO:1, or an antibody against the IL-18 receptor β subunit, which is characterized by binding specifically to a polypeptide consisting of the amino acid sequence shown by SEQ ID NO:2, is prepared.

The present invention provides progastrin-binding molecules specific for progastrin that do not bind gastrin-17(G17), gastrin-34(G34), glycine-extended gastrin-17(G17-Gly), or glycine-extended gastrin-34(G34-Gly). Further, the invention provides monoclonal antibodies (MAbs) selective for sequences at the N-terminus and the C-terminus of the gastrin precursor molecule, progastrin and the hybridomas that produce these MAbs. Also provided are panels of MAbs useful for the detection and quantitation of progastrin and gastrin hormone species in immuno-detection and quantitation assays. These assays are useful for diagnosing and monitoring a gastrin-promoted disease or condition, or for monitoring the progress of a course of therapy. The invention further provides solid phase assays including immunohistochemical (IHC) and immunofluorescence (IF) assays suitable for detection and visualization of gastrin species in solid samples, such as biopsy samples or tissue slices. The progastrin-binding molecules are useful therapeutically for passive immunization against progastrin in progastrin-promoted diseases or conditions. Also provided are surrogate reference standard (SRS) molecules that are peptide chains of from about 10 to about 35 amino acids, wherein the SRS molecule comprises at least two epitopes found in a protein of interest of greater than about 50 amino acids. Such SRS molecules are useful as standards in place of authentic proteins of interest.

The present invention relates to an integrated method for enriching and detecting rare cells in biological body fluid sample. The enriching method comprises: (a) removing plasma protein by centrifugation; (b) optionally adding a red cell lysis solution to carry out red cell lysis so as to remove the red blood cells; (c) adding immunomicrospheres or immunoadsorbent to incubate; and (d) carrying out density centrifugation based on a special cell separation medium for separating the circulating rare cells, residual red blood cells after removing red blood cells and the white blood cells combined on the immunomicrospheres. The method for detecting the enriched rare cells according to the present invention comprises combining immunohistochemistry based staining with immunofluorescence, or bicolor, tricolor or multicolor staining based on immunohistochemistry, and observing and identifying by fluorescence or ordinary optical microscope or a scanner based on microscope principle. The novel and unique method for enriching and staining has been proved to have low cost and can rapidly, effectively and high specifically enrich and quantitatively detect the rare cells in blood.

A method is provided in various embodiments for determining a profile data set for predicting the survivability of a subject with prostate cancer based on a sample from the subject, wherein the sample provides a source of RNAs. The method includes using amplification under measurement conditions that are substantially repeatable for measuring the amount of RNA corresponding to at least 1 constituent from Table 1. Alternatively, the method uses electrophoresis or immunohistochemistry for measuring the mount of protein corresponding to at least 1 constituent from Table 20. The profile data set comprises the measure of each constituent.

The present disclosure provides processes for describing and quantifying the expression of human programmed death ligand-1 (PD-L1) in tumor tissue sections as detected by immunohistochemical assay using an antibody that specifically binds to PD-L1. The results generated using these processes have a variety of experimental, diagnostic and prognostic applications.

Conjugate compositions are disclosed that include a specific-binding moiety covalently coupled to a nanoparticle through a heterobifunctional polyalkyleneglycol linker. In one embodiment, a conjugates is provided that includes a specific-binding moiety and a fluorescent nanoparticle coupled by a heterobifunctional PEG linker. Fluorescent conjugates according to the disclosure can provide exceptionally intense and stable signals for immunohistochemical and in situ hybridization assays on tissue sections and cytology samples, and enable multiplexing of such assays.

The present invention provides a method and kit for detection of two or more target molecules in a single tissue sample, such as for gene and protein dual detection in a single tissue sample. Methods comprise treating a tissue sample with a first binding moiety that specifically binds a first target molecule. Methods further comprise treating the tissue sample with a solution containing a soluble electron-rich aromatic compound prior to or concomitantly with contacting the tissue sample with a hapten-labeled binding moiety and detecting a second target molecule. In one example, the first target molecule is a protein and the second is a nucleic acid sequence, the first target molecule being detected by immunohistochemistry and the second by in situ hybridization. The disclosed method reduces background due to non-specific binding of the hapten-labeled specific binding moiety to an insoluble electron rich compound deposited near the first target molecule.

The invention discloses a preparation method of a brain tissue rapid freezing section. According to the method, acetone is used as a freezing embedding medium and a liquid nitrogen intermediate medium, so that brain tissue can be rapidly frozen , and formation of ice crystals on the brain tissue can be effectively reduced; besides, factors affecting manufacturing quality, such as temperatures during material drawing and sectioning, fixing and soring conditions after sectioning and the like, of the freezing section are optimized, and the quality of the finally prepared section is further guaranteed. According to the brain tissue rapid freezing section obtained with the method, the freezing section which has stability and can well protect antigens and the enzyme activity is provided for further immunofluorescence, immunohistochemistry and in situ hybridization tests on the brain tissue, and a pathological section meeting requirements is provided for future clinical diagnosis related to nervous system diseases and scientific researches. Particularly, molecular and biochemical index detection can be further performed on other tissue of a mouse which is not subjected to perfusion treatment in the scientific research.

The invention relates to the technical field of biology and in particular discloses an anti-silk fibroin polyclonal antibody and a preparation method thereof. The anti-silk fibroin polyclonal antibody is produced by an animal under the condtion that polypeptide with an amino acid sequence shown in SEQ ID NO:1 is taken as an immunogen. The anti-silk fibroin polyclonal antibody has the characteristics of good specificity and high purity, can be specifically conjugated with silk fibroin and is widely applicable to immunohistochemistry, immunoblotting and in vitro immunoassay. The preparation method of the anti-silk fibroin polyclonal antibody is simple in operation and is applicable to preparation of a large quantity of anti-silk fibroin polyclonal antibodies.

A sample processing system that may be configured to achieve parallel or coincidental sample processing such as histochemical processing may involve a plurality of samples arranged for coincidental movement perhaps by use of angular microscopic slide movements to cause processing activity that may include repeated elimination and reapplication of a fluidic substance perhaps through the action of capillary motion in order to refresh a microenvironment adjacent to a sample such as a biopsy or other such sample. Snap in antibody and other substances may be included to ease operator actions and to permit location specific substance applications perhaps by including single container multiple chamber multiple fluidic substance magazines, linearly disposed multiple substance source, and primary antibody cartridges. Through refreshing of a microenvironment, depletion of the microenvironment is avoided and the time necessary for slide processing may be dramatically shortened from a more common 60 to 120 minutes to perhaps less than 15 minutes so as to permit use of such a system in an intraoperative or surgical environment such as recommended by the College of American Pathologists intraoperative guidelines or the like. Patients may thus avoid a need to be subjected to an additional surgical procedure when lab results become available to see if tumors or the like were fully removed in a prior procedure.

The invention discloses a method for building an animal model with hyperuricemia-combined diabetes. The method comprises the following steps: 1) feeding an target animal with fodder containing 10% (by mass) of yeast powder; subjecting the target animal to a lavage with a 4% (by mass) adenine water solution according to the following dosage of 100 milligram of adenine per day per kilogram of body weight until the blood uric acid level of the target animal starts falling; 2) halving the dosage of the adenine water solution in step 1), subjecting the target animal to a subcutaneous injection with an oteracil potassium emulsion twice a day according to the following dosage of 100 milligram of oteracil potassium emulsion per day per kilogram of body weight until the target animal shows clinical symptoms of diabetes, and then obtaining the animal model with the hyperuricemia-combined diabetes. The method builds the animal model with the hyperuricemia-combined diabetes for the first time, the animal model has typical disease symptoms, the quantity index has statistical significance, and studies such as glucose clamp test and immunohistochemistry also prove that the modeling is successful. Therefore, the method for building the animal model with the hyperuricemia-combined diabetes is a reliable and practical technique.

An in vitro method of making a homogenous cell block for use as a positive control for biomarkers in immunohistochemistry experiments, such slide scanning and image analysis. The homogenous cell block is produced using a three layered vertical apparatus to create an evenly distributed suspension of FFPE cells, wherein the cells are mixed with 3% agarose while still rotating within the apparatus's middle layer. The injection of the cell mixture into a mold creates a homogeneous cell block where each cell, or ratio of different types of cells, is evenly distributed. The cell mixture within the cell block may comprise: a mixture of the same type of cell with different genetic modifications; a mixture of the same type of cell with different protein or nucleic acids expression; and a mixture of different types of cells with different genetic backgrounds, and / or different expression level of genes and / or proteins.

A sample processing system that may be configured to achieve parallel or coincidental sample processing such as histochemical processing may involve a plurality of samples arranged for coincidental movement perhaps by use of angular microscopic slide movements to cause processing activity that may include repeated elimination and reapplication of a fluidic substance perhaps through the action of capillary motion in order to refresh a microenvironment adjacent to a sample such as a biopsy or other such sample. Snap in antibody and other substances may be included to ease operator actions and to permit location specific substance applications perhaps by including single container multiple chamber multiple fluidic substance magazines, linearly disposed multiple substance source, and primary antibody cartridges. Through refreshing of a microenvironment, depletion of the microenvironment is avoided and the time necessary for slide processing may be dramatically shortened from a more common 60 to 120 minutes to perhaps less than 15 minutes so as to permit use of such a system in an intraoperative or surgical environment such as recommended by the College of American Pathologists intraoperative guidelines or the like. Patients may thus avoid a need to be subjected to an additional surgical procedure when lab results become available to see if tumors or the like were fully removed in a prior procedure.

The invention discloses a mouse monoclonal antibody against a human Siglec-15 protein. The specificity is high, the performance is stable and the titer is high. The invention further relates to application of the monoclonal antibody to preparation of immunohistochemical detection tools for the Siglec-15 protein. According to the mouse monoclonal antibody against the human Siglec-15 protein, an immunohistochemistry method is used for detecting expression of Siglec-15 on the tumor cell surface for selection, the curative effect and the prognosis of therapeutic schedules of malignant tumors. Theinvention further relates to the application of the monoclonal antibody to preparation of antibody drugs for immunotherapy of the malignant tumors. The important application value is achieved. The adopted antibody drugs can be full-length or partial sections (Fab, F(ab)'2 or ScFv) of an Siglec-15 monoclonal antibody or combined use of the Siglec-15 monoclonal antibody and a PD-L1 monoclonal antibody. The malignant tumors mainly include colorectal cancer, endometrial cancer, thyroid cancer, bladder cancer, kidney cancer, lung cancer, liver cancer and the like.