39 results about "Immunostaining" patented technology

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.

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.

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.

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.

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.

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 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.

This focus state evaluation method, which is for evaluating the focus state of a stain image obtained by an immunostaining method, includes: a step (step S13) for extracting a stained region from a stain image of a sample, the stain image being obtained by an immunostaining method; a step (steps S15-S17) for setting an evaluation region, including the stained region, for the stain image, and performing mask processing for excluding an exclusion region, which is a region other than the evaluation region, to acquire an image to be processed; and a step (step S18) for examining a focus state with respect to the evaluation region in the image to be processed, and acquiring a focus evaluation value indicating the focus state of the stained region. Due to the foregoing, the focus state of the stained region in the stain image can be accurately evaluated.

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.

An application of a reagent for detecting the expression level of a molecular marker in a biological sample in preparing a product for diagnosing lung cancer in a subject, wherein the molecular marker includes a combination of any two or three of COL1A1, PCDH18 and TUBB4A. The reagent is used for detecting mRNA level by a method of real-time fluorescent quantitative reverse transcription polymerase chain reaction, polymerase chain reaction, nuclease protection assay, in situ hybridisation, nucleic acid microarray, serial analysis of gene expression, Western blot, enzyme-linked immunosorbent assay, radioimmunoassay, radioimmunodiffusion, immunoelectrophoresis, tissue immunostaining, immunoprecipitation, complement fixation, fluorescence-activated cell sorting, mass analysis or protein microarray. The reagent comprises a primer, a probe and an antibody and the product comprises a kit, nucleic acid membrane strip, a preparation and a chip. The lung cancer can be lung squamous cell carcinoma.

A supramolecular histochemistry system for staining, the system includes chaotropic ionsand chaotropic ion complexing agents that act as molecular hosts to accept chaotropic ions as a molecular guest. The chaotropic ion promotes diffusion of a probe, such as an antibody into and within a tissue sample with subsequent binding of the guest to the host, or its dilution, to promote association of the antibody with a target antigen and production of an immunostaining or histochemical signal. Alternatively, small-molecule fluorescent probes can act as a molecular guest and complexed by supramolecular host to facilitate its deep penetration. A method for performing histology employs the supramolecular system with a deep homogeneous histochemistry. The method is relatively rapid, scalable, automatable, and cost effective.

The present application relates to the technical field of pathological tissue imaging, and discloses a pathological imaging system and an imaging method based on fluorescence immunotechnology, including: an immunodetection kit: including biomarkers and fluorescent materials, for clinical pathological tissue processing and staining; a polymer cell sieve: for screening and embedding clinical pathological tissues; a photographing platform: for photographing the clinically pathological tissues after staining to form pathological pictures; an AI film reading system: for reading the pathological pictures and completing the annotation of the pathological pictures. It can reduce labor and at the same time reduce the dependence on the "film reading experience" of pathologists; it does not use the paraffin embedding technique, thus shortening the time; fluorescence immunostaining improves the accuracy of AI pathological film reading; multiple biomarkers can be stained at one time, reducing the usage amount of tissue samples.