24 results about "Immunostaining" patented technology

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.

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

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.

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.

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.

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 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.

The invention relates to the technical field of immunofluorescence imaging, in particular to an automatic high-flux immunostaining sample preparation system and a use method thereof, and the system comprises an upper computer, a plurality of groups of adapters, a peristaltic pump a, a peristaltic pump b, a plurality of communication modules, a channel switching valve a, a channel switching valve b and a channel switching valve c; the upper computer is connected to the peristaltic pump a, the peristaltic pump b, the channel switching valve a, the channel switching valve b and the channel switching valve c through the communication module, the peristaltic pump b is connected with a waste liquid bottle, the peristaltic pump a is connected to the sample supply assembly through the channel switching valve c, sample preparation liquid is loaded in the sample supply assembly, and the adapters are arranged between the channel switching valve a and the channel switching valve b. The channel switching valve a is connected to the peristaltic pump a, the channel switching valve b is connected to the peristaltic pump b, and the channel switching valve a is connected to the waste liquid bottle. According to the invention, the flux of automatic immunostaining sample preparation is improved, automatic high-flux preparation of samples is realized, and the working efficiency is improved.

A supramolecular histochemistry system for staining includes a chaotropic ion and a chaotropic ion complexing agent that acts as a molecular host and accepts the chaotropic ion as a molecular guest. The chaotropic ion facilitates the diffusion of a probe, such as an antibody, into and within a tissue sample, binding the probe to the molecular host or diluting the probe, thereby promoting the association of the antibody with a target antigen and generating an immunostaining or histochemistry signal. Alternatively, a small fluorescent probe serves as the molecular guest and is complexed by the supramolecular host to facilitate deep penetration. A method for performing histology employs the supramolecular system with deep, uniform histochemistry. Such a method is relatively rapid, scalable, automatable, and cost-effective.

The invention discloses an application of FXYD1 protein in sperm centromere detection. The amino acid sequence of the FXYD1 protein is as shown in SEQ ID NO. 1. By identifying an antibody of the FXYD1 protein, the expression position of the FXYD1 protein on the sperm is marked, and the number and position of centromere in the sperm are analyzed by combining immunostaining and microscope observation. The method can effectively evaluate the sperm centromere structure state, including existence, number and position form of centromere; the related expression of the sperm FXYD1 protein can also be evaluated. The human sperm quality evaluation method provided by the invention is helpful for objectively judging sperm functions, embryonic development after fertilization, physiological and pathological states related to male fertility and prognosis results of the physiological and pathological states, and has important transformation and popularization values and wide application prospects in the fields of reproductive biology and reproductive medicine.

Endonuclease V Immunostaining Assay (EndoVIA) is provided herein as the first approach for quantifying and visualizing the landscape of Adenosine-to-Inosine (A-to-I) edited RNAs in situ. EndoVIA provides rapid quantification of overall inosine abundance and allows cell-to-cell comparison of A-to-I editing levels without the need for RNA sequencing. EndoVIA contributes valuable new insights into the dynamic interplay between A-to-I editing and subcellular localization that are undetectable with currently available approaches.

The invention discloses an immunofluorescence staining method and device for biological tissues. The biological tissue immunofluorescence staining method comprises the steps of tissue fixation, tissue degreasing and immunostaining. The tissue degreasing comprises the step of degreasing the tissue by adopting a small-micelle zwitterionic surfactant; the immunostaining comprises the step of carrying out immunostaining under a low-voltage condition of 10-100V. According to the immunofluorescence staining method, the small-micelle zwitterionic surfactant is combined with low-voltage current for immunostaining, rapid and uniform staining of large-volume tissues such as mouse brains can be realized on the premise of not causing obvious tissue damage or expansion, and a fine structure is clearly displayed.

The application discloses a kind of gastric cancer biopsy sample MHC-II expression detection immunostaining reagent, belong to the field of bioengineering.The application establishes a kind of immunohistochemical staining section preparation method, specifically including step one early treatment: tissue section baking, dewaxing and rehydration, and subsequent addition antigen solution repair, addition endogenous enzyme inhibitor incubation, addition blocking agent blocking etc.;Step two antibody incubation: addition first second antibody incubation;Step three late treatment: including staining, re-staining, reverse blue and mounting etc..With mixed buffer as endogenous enzyme inhibitor, effectively reduce the background staining intensity, improve the sensitivity and specificity of staining;At the same time, the standard of interpretation of staining results is established, which confirms that MHC-II expression is significantly related to pathological remission of gastric cancer patients, providing an evaluation method for pathological remission of gastric cancer patients.

The present invention provides a staining condition evaluation method for evaluating staining conditions of a specimen by an immunostaining method, the method comprising: a step (step S11) for imparting a primary antibody to a test specimen under one staining condition; a step (step S12) for imparting a secondary antibody and a developer to the test specimen after 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 imparting a secondary antibody and a developer to the test specimen after the step S14; and a step (step S17) for determining, after the step S16, whether or not the one staining condition is appropriate by checking the presence or absence of stained cells in the test specimen. As a result, it is possible to easily determine whether or not the one dyeing condition in step S11 is appropriate.

Endonuclease V Immunostaining Assay (EndoVIA) is provided herein as the first approach for quantifying and visualizing the landscape of Adenosine-to-Inosine (A-to-I) edited RNAs in situ. EndoVIA provides rapid quantification of overall inosine abundance and allows cell-to-cell comparison of A-to-I editing levels without the need for RNA sequencing. EndoVIA contributes valuable new insights into the dynamic interplay between A-to-I editing and subcellular localization that are undetectable with currently available approaches.

The application discloses a monoclonal antibody capable of specifically recognizing a baculovirus gp64 protein and an application thereof, the monoclonal antibody is secreted by a hybridoma cell strain 7B5-2, the hybridoma cell strain 7B5-2 has a preservation number of CGMCC No. 46131 and a preservation date of November 14, 2024. The WB experiment proves that the monoclonal antibody secreted by the hybridoma cell 7B5-2 can specifically recognize the baculovirus gp64 protein, and good application effects are achieved in an immunostaining method and a direct immunofluorescence method. The monoclonal antibody has no cross reaction with other viruses, has higher sensitivity and specificity compared with a gp64 antibody product on the market, and has higher batch precision, and can be applied to various baculovirus titer determination methods, further develops a baculovirus titer determination application scene, and provides a better antibody selection for research and development of baculovirus related diagnosis and treatment products.

Provided is a technique useful for predicting the prognosis of a patient suffering from follicular lymphoma. The present invention is a method for quantifying neoplastic follicular regulatory T cells and neoplastic follicular killer T cells from among T cells collected from a subject for predicting the prognosis of follicular lymphoma, the method comprising: a flow cytometry step in which the follicular regulatory T cells and neoplastic follicular killer T cells are quantified using an anti-CD4 antibody, an anti-CD8a antibody, an anti-PD-1 antibody, an anti-CD25 antibody, and an anti-TIM-3 antibody; detecting and counting neoplastic follicular regulatory T cells or neoplastic follicular killer T cells in the T cells collected from the subject by a flow cytometry method; the present invention relates to a method for detecting and counting neoplastic follicular regulatory T cells or neoplastic follicular killer T cells from among T cells collected from a subject by a multiple immunoassay 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, or a multiple immunostaining step in which neoplastic follicular regulatory T cells or neoplastic follicular killer T cells are detected and counted by a multiple immunoassay using an anti-CD4 antibody, an anti-CD8 antibody, an anti-PD-1 antibody, an anti-FOXP3 antibody, an anti-TCF-1 antibody,

The invention provides a specific M2-TAMs immunodiagnosis marker related to lung adenocarcinoma and application thereof. The technical key point is that SPAG4 is applied to preparation of a tumor diagnosis product as a diagnosis marker. In the early stage, a WGCNA algorithm is adopted, transcriptome data of lung adenocarcinoma patients in TCGA and GEO databases and M2 type macrophage distribution proportion data analyzed by CIBERSORT are integrated, and an M2 type macrophage polarization correlation module is successfully constructed. Through differential gene screening, Cox regression analysis and Kaplan-Meier survival analysis, the SPAG4 is finally determined as the core regulatory gene. Experimental results show that M2 type macrophage polarization and angiogenesis can be remarkably reduced by inhibiting expression of SPAG4, so that tumor growth is inhibited. On the basis, the advantages of an unmarked metabolism living body imaging technology, a living body blood vessel imaging technology, a living body immunostaining technology and molecular epidemiological research are integrated, the regulation and control mechanism of the SPAG4 gene in the lung adenocarcinoma progress and the influence of the SPAG4 gene on macrophage polarization and blood vessel generation are focused, and a novel tumor diagnosis technology and a kit are developed; the accuracy of early screening of the lung cancer is improved, and a new target spot is provided for precise treatment.