124 results about "Immune histochemistry" patented technology

The invention discloses an exfoliative cells preserving fluid which is prepared from a pH buffering agent, an osmotic pressure maintenance agent, preservatives, a fixing agent for maintaining cellular morphology, an anticoagulant, a mucus softener, an antimicrobial reagent, a cleaning agent, a humectant and red blood cell destroying components. The components in the preserving fluid are reasonable in proportioning, and the exfoliative cells can be preserved at a long time under normal temperature, wherein the longest time can reach 2 years; mucus can be sufficiently dissolved and the red cells can be partially destroyed; a film production effect is good, cellular distribution is very even, cellular morphology is perfect, cytoplasm and cell nucleus demarcation is distinct, gradation is clear, and cytoplasm and cell nucleus transparency is very good, and the exfoliative cells preserving fluid can be simultaneously used for the HPV-DNA (human papillomavirus-deoxyribonucleic acid), Chlamydia and immunohistochemical test; the special liquid base preserving fluid used for the cytologic examination of other parts can be provided, an imported product can be completely replaced, and the cellular constituent with diagnostic significance can be sufficiently preserved; and the exfoliative cells preserving fluid is low in configuration cost and easy to popularize.

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 provides a high-specificity high-sensitivity coordinated compound anti-human-TK1 antibody prepared from an antigenic determinant composed of human cervical cancer cell TK1 monomer N-terminal 23 peptide, C-terminal 20 peptide and C-terminal 28 peptide, and application thereof in tumor diagnosis. The antigenic determinant contains the following amino acid sequences: N-terminal 23 peptide (3-25): CINLPTVLPGSPSKTRGQIQVIL, C-terminal 20 peptide (206-225) CPVPGKPGEAVAARKLFAPQ, and C-terminal 28 peptide (198-225) AGPDNKENCPVPGKPGEAVAARKLFAPQ. The invention also relates to a method for preparing the antibody prepared from the antigen. The antibody kit provided by the invention has the characteristics of high sensitivity, high specificity, low cost and the like. The treatment effect on the tumor patient is evaluated by an enhanced chemiluminescent point blotting detection method, immunohistochemical detection and the detection kit.

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 provides a high-specificity and high-sensitivity coordination combination anti-human TK1 antibody prepared from an antigenic determinant consisting of 23 peptide at an N end, 20 peptide at a C end and 28 peptide at the C end of human cervical cancer cells, and application thereof to tumor diagnosis. The antigenic determinant comprises the following amino acid sequences: 1) the 23 peptide (3-25) at the N end: CINLPTVLPGSPSKTRGQIQVIL: 2) the 20 peptide (206-225) at the C end: CPVPGKPGEAVAARKLFAPQ; and 3) the 28 peptide at the C end: AGPDNKENCPVPGKPGEAVAARKLFAPQ. The invention simultaneously provides a method applying the antigen for preparing the antibody. An antibody kit provided by the invention has the characteristics of high sensitivity, high specificity, low cost and the like, tumor rehabilitation populations are periodically monitored by an enhanced chemiluminescence dot blot detection method, immunohistochemistry detection and the detection kit, and the recurrence and transfer risk and the prognosis of the tumor rehabilitation people are evaluated.

The invention discloses a method for quantifying a tumor immune state based on radiomics, wherein the method comprises the following steps: S1, collecting clinical information, immunohistochemical characteristics, overall survival OS and disease-free survival DFS of tumor patients; S2, obtaining a CT image sample group of solid tumors; S3, randomly dividing the patients into a training set and a test set in proportion; S4, calculating the immunoscore by an immune marker, and dividing the patients into a set with high immune state and a set with low immune state; S5, segmenting an interest region, and extracting radiomics characteristics of the solid tumors in the interest region; S6, screening the radiomics characteristics related to the immune state from the radiomics characteristics in the training set, and training a prediction model; and S7, predicting the image immunoscore of the tumor patients, obtaining the image immune state, and then predicting the DFS and the OS in the patients. The immune state of the solid tumors such as colorectal cancer, lung cancer, gastric cancer and breast cancer is predicted by the CT image characteristics, and the analysis of pathological sections and immunohistochemistry is reduced or avoided.

Pap smears and HPV infection tests do not distinguish between lesions that will progress to an invasive carcinoma and those that will not. We aimed to identify epigenetic biomarkers for diagnosis and progression monitoring of premalignant lesions in cervical cancer. Hypermethylated genes were identified as potential biomarkers after validation by MSP, including GGTLA4 and ZNF516. The methylation frequency for these two genes was higher in tumor: GGTLA4 (100%) and ZNF516 (96%); than in normal samples: GGTLA4 (12%) and ZNF516 (16%). The methylation status of GGTLA4 showed a progression in methylation frequency from normal samples to invasive carcinoma. The immunohistochemical expression was lower in tumor for both: GGTLA4 (50.8%) and ZNF516 (66.2%); than in normal samples: GGTLA4 (71.2%) and ZNF516 (88.1%) (p<0.05). In conclusion, we identified methylation biomarkers for the molecular screening and characterization of cervical cancer.

The present invention discloses a mouse anti-human CD41 monoclonal antibody hybridoma cell system, a monoclonal antibody and an immunohistochemical kit and an application thereof. A traditional hybridoma preparation technology is used to obtain hybridoma cell strain which excretes anti-human CD41 molecular monoclonal antibody stably. The excreted antibody can identify the conformational epitopes of the combination of human CD41 antigen and CD61 antigen and can inhibit the aggregation effect caused by ADP, collagen, HIP2, etc. The monoclonal antibody is marked biotin directly and combined with alkali phosphatase-strep avidin working fluid, substrate solution and fast red to form the immunohistochemical kit which uses SAP method to detect megakaryocyte. The kit and the detection method provided by the present invention can shorten the detection time and avoid repeated washing, so the status of cell chipping can not occur, the detection efficiency is improved greatly and the present invention is fit for common technicians to operate.

The invention provides an immunohistochemical hematoxylin-toluidine blue double-redyeing method for melanocyte. The method comprises the following steps: (1), carrying out conventional xylene dewaxing on a slice and carrying out gradient alcohol dehydration; (2), blocking and inactivating endogenous peroxide enzyme; (3), repairing an antigen; (4), carrying out serum sealing; (5), dropwise adding primary antibodies; (6) dropwise adding secondary antibodies; (7), carrying out DAB developing; (8), carrying out hematoxylin redyeing and hydrochloric acid alcohol differentiation; (9), redyeing by a toluidine blue working solution and carrying out glacial acetic acid differentiation; (10), dehydrating to be transparent and sealing the slice. According to the invention, positive granules are still brownish yellow after redyeing, and original melanin granules in the tissue become deep green, so that distinction is obvious, the melanin granules are easily distinguished from brown granules, the developing of hematoxylin is not influenced, cell nucleuses of background cells are clearly visible, the color difference of immunohistochemical positive brownish yellow granules, deep green melanin granules and blue cell nucleus is obvious and intuitive, and distinguishing is convenient. Toluidine blue dyeing liquid is prepared simply and conveniently.

The invention relates to the biotechnology area, and the spinal metastatic cancer prognosis judgment is the key to correct determine clinical development strategy, and in the invention, for the present spinal metastatic cancer related molecular biology prognosis judgment evaluation lacking problem, it constructs the spinal metastatic cancer prognosis related molecular marker tissue chip. By screening the clinical data and determining the random data spinal metastatic cancer pathological wax block to produce the spinal metastatic cancer tissue chip; selecting a variety of molecular markers and tissue chips for immunohistochemistry or in-situ hybridization, and through image analysis and quantitative measurement to analyze the expression instance of each molecular marker in the spinal metastatic cancer pathological specimens, and to analyze the relationship between the molecular markers expression and the patients prognosis, and screening the molecular markers which are closely related to the spinal metastatic cancer prognosis. The tissue chip provides a high-throughput, large sample, rapid detection tool for screening the spinal metastatic cancer prognostic related molecular markers, and provides a theoretical basis for the rational determination of the spinal metastatic cancer development strategies.

The invention discloses a construction method and use of a model of hepatitis B virus infection in vivo. The construction method of the model of hepatitis B virus infection in vivo, which uses stem cells of the human fetal liver in anti-hepatitis B virus medicament screening, comprises the following steps: separation and cultivation of the stem cells of the human fetal liver; immunohistochemical detection of the stem cells of the human fetal liver; hepatitis B virus serum infection of the stem cells of the human fetal liver; and verification of the infection of the stem cells of the human fetal liver by the hepatitis B viruses and the propagation of the viruses. The construction method, which uses the stem cells of the human fetal liver in the model of the infection in vivo, is used to screen and evaluate anti-hepatitis B virus medicaments. The method has the advantages that: the stem cells of the human fetal liver can be infected by the anti-hepatitis B viruses and constantly secrete anti-hepatitis B viruses; the stem cells of the human fetal liver can be passed and cultured; the method is simple and convenient; and the repeatability is desirable.

The present invention relates to methods and compounds for detection of molecular targets, such as biological or chemical molecules, or molecular structures, in samples using a host of experimental schemes for detecting and visualizing such targets, e.g. immunohistochemistry (IHC), in situ hybridization (ISH), ELISA, Southern, Northern, and Western blotting, etc.

The invention provides a PET / CT (Positron Emission Tomography / Computed Tomography) macroscopical digital information and pathological microscopic information matching method. A postoperative sample is reduced into an in-vivo state and is soaked into formalin; a cross section is formed between the tracer highest-ingestion layer surface in a PET / CT image and an adjacent anatomic landmark; the sample is cut apart from the middle in the cross section direction, and is soaked into the formalin; HE (Hematoxylin-Eosin) staining pathological sections are manufactured; a paraffin block region of an immune tissue chemical staining region in each HE staining pathological section is determined; the selected paraffin block region is fixed, and the paraffin section cutting is performed; and the parameter of the tracer high-ingestion layer surface in each layer of PET / CT image and the corresponding layer immunohistochemistry expression are analyzed. The PET / CT image information and immunohistochemistry information matching accuracy is improved; a tracer high-ingestion region and a histology region are accurately registered; and after the matching, the accuracy of the PET / CT on the detection of the in-vivo oncobiology information can be verified.

The invention discloses an application of combined utilization of triptolide cisplatin in preparation of a pancreatic cancer drug against drug resistance. Experiments on two drug-resistant pancreatic cancer cells PANC-1 and MIAPaCa-2 and nude mice with inoculated with PANC-1 cells on flanks prove that the effects of synergistic tumor inhibition can be achieved for drug-resistant pancreatic cancer through the combined utilization of triptolide cisplatin. So far, no study on the synergistic inhibition of the pancreatic cancer through the combined utilization of triptolide cisplatin is reported. In the nude mouse xenografting model, the tumor growth can be significantly inhibited through the combined utilization of low doses of triptolide and cisplatin, and cannot be inhibited when only the triptolide or cisplatin is used. The results of immunohistochemistry for each group of tumor biopsies show that the HSP27 protein expression can be reduced significantly through the combined utilization of the two drugs, and the results are identical to the results on cell lines. Therefore, the triptolide and cisplatin can be used together as a sensitizing agent of anticancer drugs.

The invention discloses angstrom silver oral liquid and a preparation method and application in gastric cancer treatment thereof. The angstrom silver oral liquid is prepared from angstrom silver element powder, medicinal sugar and high-purity distilled water. The preparation method comprises the steps that the angstrom silver element powder is added to the high-purity distilled water to obtain anangstrom silver solution; dispersing treatment is conducted on the angstrom silver solution; the medicinal sugar is added to the dispersed angstrom silver solution, and dispersing treatment is conducted again; the dispersed angstrom silver solution is heated and sterilized, and the angstrom silver oral liquid is obtained after natural cooling is conducted. Angstrom silver injection has significantgastric cancer cell proliferation and migration inhibition capability. The appearance and weight of primary organs are not significantly changed after oral consecutive application of angstrom silveris conducted for 15 days, and H&E dyeing and immunohistochemical analysis prove that the expression amount of tissues structures and activated Caspase-3 of the primary organs after angstrom silver application is conducted is not significantly changed.

The invention provides a research method for applying TLR4 to curing acute kidney injury caused by sepsis, and relates to the technical field of medical treatment. The method comprises the steps of mouse septic AKI model building; animal modelling judgement; specimen collection; normal and septic AKI mouse kidney cell primary culture; TLR4 determination by means of immunohistochemistry (SP method)and semi-quantitation; Real-time PCR detection of expression change of VEGF, sFLT-1, TNF-alpha and TLR4mRNA of septic AKI mouse primary kidney cells; detection of protein level expression change of VEGF, sFLT-1, TNF-alpha and TLR4 of septic AKI mouse primary kidney cells through a Western-blot method; constructing VEGF gene eukaryotic expression plasmids and optimal silence mouse VEGF gene ShRNAexpression plasmids; experimental grouping; TLR4 cell treatment; index detection; verifying TLR4 target regulation on VEGR expression; searching for protein in interaction with the VEGF; discussing molecule mechanisms which is possibly involved in TLRs signal path adjustment of the VEGF. The method speculates that the VEGF and the TLR4 signal path are in interaction for regulating the acute kidneyinjury caused by sepsis and play a part in protection.

The invention relates to a NF-kappaB inhibitor, process for preparation, and use as antineoplastic medicine, wherein extracorporal pointing mutation method is applied twice for performing orienting mutagenesis to the codon of the No.32 and No.36 bit encoding serine to construct IkappaBalphaM super inhibitor, which is than sub-cloned from pGEM-T carrier into pAdTrack-CMV shuttle vector, then it is expressed in pronucleus and eukaryotic cells (including 293.HepG2, HeLa, HUVEC, MAD-435S cells, etc.), thus obtaining the protein (expressed NF-kappaBsuperinhibitor IkappaBalphaM).

The invention discloses a compound type biological tissue and cell chip, relating to the subjects such as molecular biology, clinical medicine, veterinary medicine, zoology, medicinal form teaching and the like. At present, single tissue chip and cell chip are under great restriction in experimental study and clinical diagnosis work, comparative study on the tissue and cell can not be implemented, the internal structure of the cell can not be observed, and the expensive antibody, time and the like are wasted; the problems are solved perfectly by adopting the compound type biological tissue and cell chip. Tissue and cell specimens are fixed, dehydrated and embedded into tissue and cell wax blocks according to a tissue specimen treatment mode; then, the materials are drawn for the second time according to the subject design, the tissue and cell specimen columns are arrayed on the same carrier before being embedded and sliced, and subsequent experiments such as immunohistochemistry and the like are carried out synchronously. The compound type biological tissue and cell chip is mainly used for performing comparative study on the tissue and cell level in a scientific research activity, observing the internal cell organelle structure of the cell and carrying out batch detection diagnosis on the clinical cytology specimens, thereby having great significance in scientific research and bringing good social and economic benefits.

The invention discloses an immunohistochemical operation method. The immunohistochemical operation method comprises the following steps of: (1) slicing and baking slices; (2) dewaxing and hydrating; (3) repairing an antigen; (4) sealing endogenous catalase; (5) dyeing; (6) dropwise adding an antibody I; (7) dropwise adding an antibody II; (8) developing, namely developing by adopting a DAB developing agent at a room temperature; and (9) sealing the slices. According to the technical scheme disclosed by the invention, a conventional operation step of dropwise adding the antibody I and the antibody II before dyeing by haematoxylin, the antibody can be easily developed after the antibody I and the antibody II are dropwise added. The immunohistochemical operation method is simple in operation steps; only an operation sequence of existing operation steps, adopted reagents and operation time are changed and the change of an existing operation effect can be realized; the antibodies are dropped after dyeing so that the size and position of tissues can be conveniently grasped. The operation steps are simple and certain technical detail problems in experiment check analysis can be effectively solved; standard operation of an experiment are easy to realize, wastes of experiment reagents can be avoided and certain errors in the experiment are also reduced as much as possible.

The invention relates to the technical field of immunohistochemical staining and particularly relates to a double-label immunohistochemical staining kit used for differential diagnosis of metastatic renal cell carcinoma. The kit comprises a DAB developing solution and a primary antibody formed by mixing a mixed solution of a PAX8 antibody and a CD10 antibody or / and an HRP enzyme-labeled secondary antibody. In order to overcome the disadvantages that the histological characteristics of the metastatic renal cell carcinoma are extremely easily confused with adrenal tumors, lung cancer, liver cancer, thyroid cancer and the like so that a definite diagnosis is not easy to make, a conventional immunohistochemical method is used for respectively detecting, more time and labor are consumed, the tissue amount of an aspiration biopsy specimen is not enough and the like, the same sensitivity and specificity when each antibody in the mixed antibody is singly stained are kept by using the method; and the double-label immunohistochemical staining kit also has the advantages that staining steps are few, the experiment time is short, the stability is high, an experiment result can be directly compared, and brought information amount is much higher than that of traditional single staining and the like, and especially has the remarkable advantages for biopsy pathological diagnosis with few specimen amounts.

This invention opens one establishing method of one fetal pancreas stem cell lines. epithelioid stem cell of fetal pancreas comes from fetal pancreas organization in normal abortion, in vitro culture can be clonned infinitely, and possesses the potential of multi-directional differentiation; after testing and analysis of morphological character, growth characteristics, immunohistochemistry characteristics, RT-PCR character, directional induced differectiation characters, chromosome karyotypes, DNA array, it is proved that this fetal pancreas stem cell is human originated fetal pancreas stem cell; this invention can provide affulent seed cells for researches on mammalian pancreas development and diabetes origination and dibetes treatment by reproducing Langerhans' islands' micro organs. It also provides an efficient and reliable method for establishing pancreas stem cell lines of creodonts and human beings.

The invention provides a preparation method of artificial cutaneous melanoma tissues, including the culture of keratinocytes and melanoma cells of human skin as well as the preparation of biological matrix material. The keratinocytes and the melanoma cells of human skin are respectively inoculated on the biological matrix material in sequence, and by the adoption of a method combining culture under liquid and air-liquid level incubation, a certain culture medium is used under the conditions of 5 percent of CO2 and asepsis at 37 DEG C to culture and construct the cutaneous melanoma tissues. Methods of histology and immunohistochemistry technology are adopted for the observation of growth situation of the malignant cutaneous melanoma tissues. The melanoma tissues closer to the microenvironment of human skin are constructed by the preparation method and the observation by the histology method shows the obvious invasive growth of the melanoma tissues and the formation of obvious tumor metastases or tumor groups on the surface of HDED and in the lumen of the dermal accessory organ, thus meeting the distribution characteristics of the malignant melanoma tissues infiltrated in the dermis of human skin and reflecting that markers of the melanoma tissues show a positive reaction.

The invention relates to a method for preparing stationary liquid for slicing and application of the stationary liquid for the slicing, and belongs to the technical field of clinical medicine application. The method comprises the steps that methyl alcohol, glutaraldehyde and diethyl ether are mixed, and PBS buffer solutions are used for attenuation; saccharose is added to the solutions to be fully mixed, a pH value is regulated, and the stationary liquid for the slicing is obtained. Tissues are treated through pretreatment, saccharose placing, embedding, slicing and washing. The method is simple in working progress and step, the operating work amount of technical personnel is reduced, the operating steps of the whole process are few, needed time is shortened, tissues free of dehydration and waxdip can be kept fresh more highly, antigens for immunohistochemistry are kept complete and excellent, more accurate diagnosis conditions can be provided, xylene and other highly-polluted poisonous reagents are not needed, environmental pollution is reduced, and the guarantee for the body health of the technical personnel is enhanced.

The invention relates to a detection method for testing a protein HOP and a heterogeneous ribosome protein C1 / C2 which are both expressed in human liver tissue, as well as reagent box used in the method. The method is characterized in that: by means of the fluorescence differential two-dimensional electrophoresis method -MALDI-TOF / TOF, the inventor manages to identify the overexpression of the two proteins on human liver cell carcinoma tumor tissue, and carries out immune histochemistry validations of Western bolt and large amount of samples over the expression levels in the human liver cell carcinoma tissue. The detection method for proteins and the reagent box can be used in the detections for clinical or research purposes.

The invention relates to the field of biotechnology, in particular to an immunohistochemical technique based pathology assistant analysis method which comprises the following steps: acquiring a communication channel connected with a processing platform through a client; uploading pathology digital images so as to obtain a cell sample interval in the pathology digital images; setting a parameter set, and searching cells matching the parameter set in the cell sample interval as target cells; counting the amounts of positive cells and negative cells in the target cells according to staining colors of the target cells respectively; obtaining a ratio of the positive cells to the target cells, and outputting the ratio to the client; evaluating the accuracy of the ratio through the client by a user, and then supplying accuracy feedback to the processing platform; and regulating the parameter set. The technical scheme of the invention has the beneficial effects that the immunohistochemical technique based pathology assistant analysis method can assist a doctor to carry out pathology analysis based on an immunohistochemical technique, also has a feedback optimization mechanism and can improve the accuracy of diagnosis while lowering the labor cost of immunohistochemical diagnosis.

The invention belongs to the biotech field, relates to an application of an Shp2 protein, and concretely relates to an application of the Shp2 protein in preparation of a liver cancer prognosis evaluation kit. The relative Shp2 protein expression level of a tumor tissue to a tissue adjacent to the tumor is determined through utilizing an immunohistochemical technology, microscope shooting and computer image processing software, and postoperative follow-up information is combined to determine that the relative Shp2 protein expression level is related with the hepatocellular carcinoma patient prognosis after the operation, so the Shp2 protein can be used for preparing a protein molecule maker for determining the liver cancer patient, and is of great guiding significance to the postoperative monitoring and sequential treatment of hepatocellular carcinoma patients.

The invention relates to the field of biotechnology, especially to a detection kit of a novel target PNC for detection of tumor invasion and application thereof. The first detection antibody of the PNC detection kit for detecting tumor invasion is a PTB or CUG-BP1 antibody. The invention describes composition and application methods of a PNC immunohistochemical detection kit and a PNC immunofluorescence detection kit used for detecting tumor invasion. Furthermore, the invention also provides a method for screening a novel anti-metastasis compound and a Chinese herbal medicine monomer thereof; and the method for screening the novel anti-metastasis compound is also applicable to screening of small molecule compounds and Chinese herbal medicine monomers thereof. According to the invention, the proportion of PNC in a tissue sample of a cancer patient obtained through paracentesis is detected by using the PNC kits, so detection is simple and convenient, is easy to operate and has good stability; the proportion of PNC in tumor cells which have been affected by the compound is detected by using the PNC kits, computer programming and analysis are cooperatively used, so high throughput screening of a novel drug is realized.