58 results about "Immunostaining" patented technology

The invention discloses a monoclonal antibody H2 specifically bound with a lung cancer marker CYFRA21-1 and application, and relates to the technical field of biological medicines. The method comprises the following steps: firstly, screening from a Tomlinson I + J phage library by utilizing a phage display technology to obtain a lung cancer marker CYFRA21-1 monoclonal antibody H2, and the antibody has a heavy chain complementary determining region and a light chain complementary determining region which are formed by novel amino acid sequences and can be specifically combined with CYFRA21-1 protein; the monoclonal antibody H2 of the lung cancer marker CYFRA21-1 has the characteristics of high affinity, high specificity, stability and the like, and the monoclonal antibody H2 of the lung cancer marker CYFRA21-1 not only can efficiently and accurately detect the concentration of CYFRA21-1 protein, but also can be used for immunostaining of lung cancer tumor cells and lung cancer tissues in pathological diagnosis, and has important significance for detecting the concentration of CYFRA21-1.

To provide a method of clearing false positive immunostaining signals and noise, to perfect incomplete staining.SOLUTION: A method for refining virtually-stained image annotations includes separating an image of a stained tissue section into an image of a counterstain and an image of immunostaining visual signals via linear transformation, to identify and segment nuclei in the image of the counterstain to generate an image of a refined counterstain with single-cell segments. The method further includes comparing immunostaining visual signals in the single-cell segments of the image of the refined counterstain to a user-defined threshold to determine true positive cells. The method further includes overlaying the image of the counterstain and the image of refined immunostaining visual signals to generate a refined virtually-stained image annotation.SELECTED DRAWING: None

A supramolecular histochemical system for staining, the system comprising chaotropic ions and a chaotropic ion complexing agent that accepts chaotropic ions as a molecular subject as a molecular guest. The chaotropic ions facilitate diffusion of probes (e.g., antibodies) into and within the tissue sample, followed by binding or dilution of the guest to the subject to facilitate binding of the antibodies to the target antigen and immunostaining or generation of histochemical signals. Alternatively, the micromolecular fluorescent probe can be used as a molecular guest to be compounded with the supramolecular subject to promote deep penetration of the supramolecular subject. A method for performing histology employs a supramolecular system with deep homogeneous histochemistry. The method is relatively fast, scalable, automated and cost effective.

The invention discloses an immunostaining reagent for MHC-II expression detection of a gastric cancer biopsy sample, and belongs to the field of bioengineering. The invention establishes a preparation method of an immunohistochemical staining section. The preparation method specifically comprises the following steps: step 1, early-stage treatment: baking a tissue section, dewaxing, rehydrating, subsequently adding an antigen solution for repairing, adding an endogenous enzyme inhibitor for incubation, adding a sealing agent for sealing and the like; step 2, antibody incubation: adding a first antibody and a second antibody for incubation; and step 3, post-treatment: dyeing, re-dyeing, bluing, sealing and the like. A mixed buffer solution is used as an endogenous enzyme inhibitor, so that the background staining intensity is effectively reduced, and the staining sensitivity and specificity are improved; meanwhile, a staining result interpretation standard is established, it is proved that MHC-II expression is remarkably related to pathological remission of the gastric cancer patient, and an evaluation method is provided for pathological remission of the gastric cancer patient.

A method for refining virtually-stained image annotations. The method includes separating an image of a stained tissue section into an image of a counterstain and an image of immunostaining visual signals via linear transformation. The method includes identifying and segmenting nuclei in the image of the counterstain to produce an image of a refined counterstain with single-cell segments. The method includes comparing immunostaining visual signals in the single-cell segments of the image of the refined counterstain to a user-defined threshold to determine true positive cells. The method includes refining the image of the immunostaining visual signals to produce an image of refined immunostaining visual signals. The refining is based on the stained tissue section protein localization or morphology and the determined true positive cells. The method includes overlaying the image of the counterstain and the image of refined immunostaining visual signals to produce a refined virtually-stained image annotation.

The invention relates to an intestinal tissue inflammation injury treatment target spot and application thereof, and belongs to the technical field of biological medicine. It is found that the level of DPF2 in intestinal tissue inflammation damage can significantly affect regeneration and repair of intestinal tissue, high-level DPF2 can promote intestinal tissue inflammation damage, and after the DPF2 gene is knocked out, intestinal tissue inflammation damage can be significantly reduced. Clinical intestinal tissues are subjected to DPF2 immunostaining, clinical cases are divided into a high-inflammation injury group and a low-inflammation injury group, the high-inflammation injury group shows a higher DPF2 level, it is indicated that DPF2 can indeed serve as a new index for evaluating and diagnosing intestinal tissue inflammation injury, and the DPF2 has a good clinical application prospect.

The application discloses a use of MHC-II expression in prediction of curative effect of gastric cancer immunotherapy and belongs to the field of bioengineering. Specifically, the application comprises the following steps: detecting MHC-II expression in a gastric cancer biopsy sample; baking, dewaxing and gradient alcohol treating the gastric cancer biopsy sample slice; carrying out antigen repairing and blocking treatment; then adding diluted MHC Class II antibody for incubation, and then adding immunopotentiator and enhanced enzyme-labeled antibody for incubation; finally, completing immunostaining through DAB color development, staining liquid staining and counter blue treatment, mounting and observing. The MHC-II expression signal is clear and distinguishable, the staining effect is good, and through negative and positive expression interpretation and pathological remission evaluation, it is confirmed that the MHC-II expression is significantly related to pathological remission of a gastric cancer patient and plays an important role in prediction of curative effect of gastric cancer immunotherapy.

Disclosed is a method for detecting circulating tumor cells containing defective APC protein, comprising immunostaining circulating tumor cells in the blood using a labeled antibody that binds to both wild-type and defective APC proteins, and a labeled antibody that binds to wild-type but not defective APC proteins, and measuring the signal derived from the label.

The present invention relates to a method for the in vitro characterization and quantification of proliferative cells in an epidermal fragment, comprising the following steps: 1) Culturing the epidermal fragment on a support, 2) Detaching the epidermal fragment from the support, 3) Performing immunostaining on the epidermal fragment, 4) Acquiring images of the basal layer of the epidermal fragment over its entire surface, 5) Reconstructing an image representing the entire basal layer of the epidermal fragment, 6) Selecting the immunostained nuclei of the proliferative cells by image processing, 7) Quantifying cell division by counting the nuclei through image analysis. The present invention also relates to using the method to evaluate the effect of a product or active ingredient on epidermal cell regeneration. Figure 1

The present invention discloses a monoclonal antibody H2 that specifically binds to the lung cancer marker CYFRA21-1 and its application, relating to the field of biomedicine technology. First, using phage display technology, a lung cancer marker CYFRA21-1 monoclonal antibody H2 is obtained by panning from the Tomlinson I+J phage library. The antibody has a heavy chain complementary determining region and a light chain complementary determining region composed of novel amino acid sequences, and can specifically bind to the CYFRA21-1 protein. The binding has the characteristics of high affinity, high specificity and stability. The monoclonal antibody H2 for the lung cancer marker CYFRA21-1 of the present invention can not only efficiently and accurately detect the concentration of CYFRA21-1 protein, but also can be used for immunostaining of lung cancer tumor cells and lung cancer tissues in pathological diagnosis, which is of great significance for detecting the concentration of CYFRA21-1.

The present invention provides a method of treating a patient diagnosed with Myxoid / Round cell liposarcoma with an anti-GPC3 therapeutic agent. The present invention also relates to quantification of GPC3 expression in tissue samples of patients diagnosed with Myxoid / Round cell liposarcoma by an immunostaining assay and identification of GPC3 expression levels that correlate with selection of patients for administering the anti-GPC3 therapeutic agent.

In order to enable use of novel immunostaining for a pathological diagnosis and research, the invention provides a sample exchange chamber including: a container 2 into which a substrate on which a sample is placed can be introduced; a specific solution inlet 3, which is a staining mechanism that stains the sample; a cleaning liquid inlet 4, which is a cleaning mechanism that cleans the sample; an evacuation port 5, which is an evacuation mechanism that evacuates the container; a drain port 6; and a sterilization mechanism 7 that sterilizes the sample and inside of the container.

To easily determine suitability of a staining condition.SOLUTION: A staining condition evaluation method for evaluating a staining condition of a specimen by an immunostaining method includes: a step (step S11) for applying a primary antibody to a test specimen under one staining condition; a step (step S12) for applying a secondary antibody and a color developing agent to the test specimen after the step S11; a step (step S14) for inactivating or removing the primary antibody on the test specimen after the step S12; a step (step S16) for applying the secondary antibody and the color developing agent to the test specimen after the step S14; and a step (step S17) for determining suitability of the one staining condition by checking whether there is a stained cell in the test specimen after the step S16. Thus, the suitability of the one staining condition in the step S11 can be easily determined.SELECTED DRAWING: Figure 1

An image processing using a plurality of specimen images obtained by successively applying a plurality of types of staining to a specimen to be evaluated and imaging the specimen after staining for at least two types of staining is performed. The image processing comprises: extracting an inner region corresponding to inward of a cell membrane of a single cell in the specimen based on at least one of the specimen images; specifying a cell region corresponding to an individual cell included in the specimen by expanding the inner region outwardly; and performing image cytometry for the cell region based on the specimen images. In analyzing a pathological specimen on a cell-by-cell basis, it is possible to deal with multiple immunostaining and specify the positions of individual cells and evaluate each cell separately even when a cell density in the specimen is high.

The present invention belongs to the field of 3D printing technology and cell culture technology. Specifically, the present invention relates to a device and method for rapid preparation of cell patterned co-culture based on 3D printing. The present application provides a microdevice technology that can be rapidly prepared using 3D printing for cell culture and cell patterned co-culture, and the device can be easily compatible with existing cell biotechnology, such as microscopic imaging, cell immunostaining, etc. It can easily achieve patterning of multiple cells, and can perform microscopic imaging analysis and immunohistochemical analysis on cells to obtain quantitative results. It provides a device and method for scientific researchers to achieve co-culture of multiple cells more quickly and conveniently.

A method and system are disclosed for differentiating non-human primate induced pluripotent stem cells (NHP iPSCs) into hepatocyte-like cells through a defined, multi-stage protocol. The method includes sequentially exposing NHP iPSCs to six distinct cell differentiation media over an approximate eighteen-day period. Each medium comprises defined combinations of growth factors, small molecules, and inhibitors that direct progressive lineage specification through anterior primitive streak, definitive endoderm, posterior foregut, liver bud progenitors, hepatoblasts, and hepatocyte-like cells. Concentration ranges and exposure durations are provided for each stage, ensuring reproducibility and reducing heterogeneity across cell populations. In some embodiments, treatment with a transforming growth factor beta (TGF-β ) receptor I inhibitor during early hepatoblast specification enhances lineage fidelity. The resulting hepatocyte-like cells are validated by immunostaining and western blotting for hepatic markers. Applications include preclinical drug metabolism, hepatotoxicity assessment, and in vitro disease modeling using non-human primate–derived systems.

A solution transfer system for tissue sample incubation. The system includes cartridges, each including a sample chamber and openings disposed on its surface. The sample chamber may hold a tissue sample. The system further includes a flow plate assembly having a main channel, and a main inlet, and a main outlet fluidly coupled to it. The flow plate assembly further includes hollow bodies, each one configured to contain a cartridge. Each hollow body further includes an inlet interface fluidly coupled to the first opening and the main channel and an outlet interface fluidly coupled to the second opening and the main channel. Each hollow body further includes valves at the inlet and outlet interfaces. The system further includes a control mechanism operatively coupled to the valves, configured to actuate each valve individually.

The application discloses a kind of based on non-diffracting airy beam intrahepatic blood vessel three-dimensional imaging method, belong to intrahepatic blood vessel three-dimensional imaging technical field, solve the problem of insufficient accuracy and detection sensitivity of existing detection method, method includes by liver transplantation or biopsy surgery obtains liver tissue, liver tissue is fixed using 4%PFA processing, liver tissue is bleached using H2O2Solution, antigen repair is carried out to liver tissue using FLASH solution, using goat serum blocking solution to the liver tissue after antigen repair blocking treatment;Liver tissue immunostaining treatment, liver tissue dehydration treatment, based on non-diffracting airy beam and high-throughput microscope to the liver tissue after processing imaging, obtain the three-dimensional imaging picture of liver tissue;The application is immunofluorescence staining and transparent treatment to liver tissue, so that laser can smoothly penetrate tissue and carry out three-dimensional imaging, can realize single wide-range image capture.

A method of producing oligodendrocytes recapitulates the major steps of oligodendrocyte differentiation in a significantly shorter time than the 75-95 days required for the previous protocol. The cannabinoid receptor agonist, WIN55212-2, induces NSC differentiation and promotes OLIG2+OPC induction. WIN-derived OLIG2+ progenitor cells differentiate into PDGFR α+ OPCs, which are highly migratory, can further differentiate into mature OLs with strong myelination ability. When WIN-derived OPCs were transplanted to the lesion site, the motor ability of spinal cord injury (SCI) mice was significantly improved from the second week after transplantation. Immunostaining results showed that WIN-derived OPCs could differentiate into mature OLs and myelinate injured axons. The method and product have wide applications in the field of cellular therapy, especially for spinal cord injury with remarkable therapeutic effects.

The present invention relates to a novel anti-PTK7, protein tyrosine kinase 7, antibody and derivatives thereof. The antibody and / or derivatives can be used for therapy and / or imaging, diagnosis and / or immunostaining.

The invention relates to an in-situ binding method for evaluating binding of Bt toxin Cry1Ac and midgut epithelial cells of cotton bollworm larvae. The method comprises the following steps: incubating and culturing midgut tissues of larvae of lepidoptera pests and a Bt toxin-containing Sf-900 II cell culture solution, carrying out immunostaining, observing and photographing through a laser confocal microscope, and finally carrying out quantitative processing on a fluorescence image by utilizing ImageJ software so as to calculate an average unit area fluorescence signal value. And further evaluating the difference between the binding amounts of the midgut epithelial cells and the Bt toxins of the larvae of different strains. The method can intuitively and accurately observe and evaluate the binding condition of the Bt toxin and the midgut epithelial cells on the premise of retaining the original structure and physiological environment of the midgut tissue of the larvae, provides direct data support for Bt resistance research, assists in analyzing the pest Bt resistance mechanism, has important significance in formulating resistance early warning and treatment strategies, and has a wide application prospect. The method is suitable for the biotechnical fields of Bt toxin insecticidal mechanism research, lepidoptera pest drug resistance mechanism exploration and the like.

An object of the present invention is to provide a matured cultured cell sheet comprising cardiomyocytes without using a specific drug and a method for producing the same. Another object of the present invention is to provide a method for evaluating a compound or drug using the cultured cell sheet and a method for evaluating the quality of the cultured cell sheet comprising cardiomyocytes. The cultured cell sheet of the present invention is a cultured cell sheet comprising cardiomyocytes, wherein the cardiomyocytes are arranged with alignment, and a degree of alignment determined according to the following formula (1) is 23% or more: degree of alignment (%)=(number of rod-shaped structures included within ±15° of mode) / (total number of rod-shaped structures)×100 (1), wherein in an image obtained by microscopy of the cultured cell sheet immunostained with an anti-α-actinin antibody, where a screen horizontal direction is regarded as 0°, a degree of alignment is a frequency of rod-shaped structures included within ±15° of the mode of angles which are measured in a longitudinal direction of the rod-shaped structures detected with an α-actinin antibody within a measurement range of 71.6 μm×71.6 μm.

Herein is disclosed amphiphilic small molecules with different hydrophobic and hydrophilic fragments that show high M binding affinity to both Aβ plaques and oligomers, and selectively binding Aβ oligomers. These amphiphilic compounds can also label the Aβ species in the brain sections of transgenic AD mice, as shown by immunostaining with an Aβ antibody. Certain amphiphilic compounds were found to alleviate the Cu2+-Aβ induced toxicity in cell viability assays. Additionally, two compounds, ZY-15-MT and ZY-15-OMe, were found to disrupt the interactions between Aβ oligomers and human neuroblastoma SH-SY5Y cell membranes. These studies show compounds with amphiphilic properties that target Aβ oligomers and modulate the Aβ oligomer-cell membrane interactions can be effective as small molecule AD therapeutics. Also, the disclosed amphiphilic dicyanomethylene compounds that can emit in the near-infrared region and chelate copper can be used as early diagnostic agents for AD.

The invention relates to a high-fidelity expansion microscopic imaging method of a nuclear pore complex, which is characterized in that the signal intensity and the dyeing coverage are obviously enhanced through the immunostaining conditions of 37 DEG C temperature and prolonged incubation time, and the permeability of an antibody in iterative expansion gel is improved; by adopting a secondary anchoring cross-linking condition that 2.8% paraformaldehyde and 10% acrylamide are proportioned, under the condition that the expansion rate is not reduced, the structural heterogeneity index is further reduced, and the marking integrity is improved; through treatment of 92.5 mM SDS under the denaturation condition of 85 DEG C for 1.5 h, full denaturation can be achieved, and deformation and signal loss caused by protein breakage can be reduced. During imaging, the signal-to-noise ratio of fluorescence after single expansion is improved by using a water immersion objective lens, so that a clear annular NPC structure can be obtained by using a confocal microscope; therefore, the image fidelity and the signal-to-noise ratio are remarkably improved, and a technical basis is provided for subsequently realizing near-molecule-scale NPC imaging under a confocal microscope.

The application discloses a surface fluorescence chromatography resin embedding method suitable for immunostaining samples, and comprises the following steps: S1, biological tissue sample pretreatment; S2, gradient ethanol dehydration and tissue blackening; S3, gradient resin permeation; S4, acidification treatment; and S5, resin polymerization molding. The application also comprises a biological tissue sample imaging method, which comprises the step of imaging the biological tissue sample obtained by the embedding method in a fMOST system sodium carbonate solution environment. The application temporarily quenches the pH-sensitive chemical probe by adding HAc; in combination with the fMOST imaging system, the reactivation is realized in the alkaline solution environment, the blackening chemical chromatography high-precision imaging is realized by reactivating the sample imaging surface, and the immunostaining sample has the advantages of high fluorescence retention rate.

A region selection assistance method comprises: a step (step S11) for dividing a stained image of a specimen obtained by an immunostaining method into a plurality of divided regions; a step (step S12) in which, for each of the plurality of divided regions, one display color is assigned from a display color group including a plurality of colors having different visibility, such that the visibility becomes higher as the aggregation of the stained cells in the divided regions becomes higher (step S12); a step (step S13) for generating a selected auxiliary image in which each of the plurality of divided regions is filled with the display color assigned in step S12; and displaying the selected auxiliary image on a screen (step S14). As a result, an operator can easily select a region suitable for analysis in the dyed image as an analysis region.

The present invention addresses the problem of providing: a mature cultured cell sheet comprising cardiac muscle cells without using a specific drug; and a method for producing the mature cultured cell sheet. Also provided are a method for evaluating a compound or a drug using the cultured cell sheet, and a method for evaluating the quality of a cultured cell sheet comprising myocardial cells. The cultured cell sheet according to the present invention is a cultured cell sheet comprising myocardial cells, the myocardial cells being arranged so as to have orientation, and the cultured cell sheet being obtained by immunostaining the cultured cell sheet with an anti-alpha-actin antibody and observing the same with a microscope, in an image obtained by subjecting the cultured cell sheet to immunostaining with the anti-alpha-actin antibody. When the angle in the longitudinal direction of the rod-like structure, which is detected by an alpha-actin antibody, is measured within the measurement range (71.6 [mu] m * 71.6 [mu] m) when the image horizontal direction is set as 0 degree, and the degree of orientation, which is obtained by formula (1), is 23% or more, said degree of orientation being the frequency of the rod-like structure contained within the mode + / -15 degrees. Degree of orientation (%) = (number of rod-shaped structures included within mode + / -15 degrees) / (total number of rod-shaped structures) * 100 (1)

The present invention provides a slide for precision management, which can estimate the presence or absence of an abnormality in an automatic immunostaining device and can estimate the cause of the abnormality when the abnormality has occurred. The slide for precision management is characterized by having a spot (antigen (-)) that does not contain an antigen, a spot (fixed (-) / antigen (+)) that contains an antigen without applying histological immobilization, and a spot (fixed (+) / antigen (+)) that contains an antigen with applying histological immobilization.

The present invention provides a technology useful for predicting prognosis in a follicular lymphoma patient. The present invention is a method for quantifying neoplastic follicular regulatory T cells and neoplastic follicular cytotoxic T cells in T cells collected from a subject, in order to determine the prognosis of follicular lymphoma. The method comprises: a flow cytometry step in which neoplastic follicular regulatory T cells and neoplastic follicular cytotoxic T cells in T cells collected from a subject are detected and counted with flow cytometry using an anti-CD4 antibody, an anti-CD8 antibody, an anti-PD-1 antibody, an anti-CD25 antibody, and an anti-TIM-3 antibody; or a multiplex immunostaining step in which neoplastic follicular regulatory T cells and neoplastic follicular cytotoxic T cells in T cells collected from a subject are detected and counted with multiplex immunostaining using an anti-CD4 antibody, an anti-CD8 antibody, an anti-PD -1 antibody, an anti-FOXP3 antibody, an anti-TCF-1 antibody, and an anti-Granzyme K antibody.

The invention discloses a tissue transparentizing method and an immunostaining method for esophagus of a multi-stage canceration mouse. The tissue transparentizing method comprises a whole tissue immunostaining step, a transparentizing step, a three-dimensional imaging step, a histological staging step and an image analysis step, wherein the whole tissue immunostaining step comprises the step of carrying out whole tissue immunostaining on a mouse esophagus sample by adopting an antibody; and a histological staging step, including de-transparentizing the three-dimensional imaging sample, and then performing pathological staining to judge the pathological staging of the sample. According to the application, a mouse esophageal whole tissue immunostaining method and a post-transparentizing esophageal original tissue pathological staging method are researched and developed for the first time, and a basis is provided for researching three-dimensional space characteristics of multi-stage lesions of mouse esophageal squamous carcinoma; the invention constructs a set of tissue transparentizing technology flow to realize microenvironment three-dimensional characteristic analysis in the mouse esophageal squamous cell carcinoma progress process.