83 results about "Antigen retrieval" patented technology

The present invention concerns a method and apparatus for automatic staining or other processing of a biological sample on a slide, perhaps robotically, by applying predetermined amounts of reagents in a predetermined sequence according to a processing protocol, the processing including pre-treatment steps, under the control of an adaptive processing control system. Further provided is a method of antigen retrieval, the method comprising contacting, within a heated processing tank of an automated staining apparatus, at least one biological sample with at least one composition for antigen retrieval comprising at least one chelating agent, at least one detergent and at least 50% by volume of at least one liquid that facilitates antigen retrieval.

A method is disclosed for reversing fixation-induced cross-linking in tissue specimens that have been preserved for histological examination. The method involves placing the fixed tissue in a liquid under elevated temperature and pressure conditions that are sufficient to reverse the fixation-induced cross-linking, restore antigenicity to proteins, and permit improved molecular and proteomic analysis of the preserved tissue specimen. Methods are also disclosed for processing tissues for histological examination under elevated pressure conditions that enhance the perfusion of liquid reagents into the tissue and reduce overall processing times.

The present invention concerns a method and apparatus for automatic staining or other processing of a biological sample on a slide, perhaps robotically, by applying predetermined amounts of reagents in a predetermined sequence according to a processing protocol, the processing including pre-treatment steps, under the control of an adaptive processing control system. Further provided is a method of antigen retrieval, the method comprising contacting, within a heated processing tank of an automated staining apparatus, at least one biological sample with at least one composition for antigen retrieval comprising at least one chelating agent, at least one detergent and at least 50% by volume of at least one liquid that facilitates antigen retrieval.

Novel methods of probing multiple targets in a biological sample are provide whereby the targets are DNA, RNA and protein. The method comprises subjecting the sample to an in situ hybridization reaction using a labeled nucleic acid probe that binds an RNA target, observing a signal, and optionally removing the signal. The method further comprises an antigen retrieval protocol, observing a signal, removing the signal, and optionally applying a protease treatment to access the sample's DNA targets by subjecting the sample to an in situ hybridization reaction using a labeled nucleic acid probe, observing a signal from the labeled DNA targets, and optionally removing the signal.

The present invention relates to the field of cancer. More specifically, the present invention provides methods and compositions useful for diagnosing or predicting cancer in a patient. In one embodiment, a method for identifying a patient as having cancer comprises the steps of (a) providing a formalin-fixed, paraffin-embedded or fresh frozen sample of patient tissue; (b) steaming the sample in antigen retrieval buffer; (c) incubating the sample in hydrochloric acid (HCl); (d) incubating the sample with an affinity reagent specific for 5hmC under conditions to form a complex between the affinity reagent and 5-hydroxymethylcytosine (5hmC) present in the sample; (e) detecting the complexes formed between 5hmC and the affinity reagent with secondary detection reagents; (f) quantifying 5hmC levels; and (g) identifying the patient as having cancer if the 5hmC levels in the sample are reduced as compared to a control.

Solutions exhibiting little or no evaporative loss at elevated temperatures, i.e., in excess of 100° C., are employed in place of conventional aqueous-based antigen retrieval solutions.

The invention discloses a multi-sample immunohistochemical detection board, and also discloses a high-throughput immunohistochemical detection method with the detection board. The method comprises the following steps of: (1) preparing a tissue section to be detected; (2) flatly putting the tissue section to be detected in a step (1) in a transverse groove (2) after dewaxing, hydrating, carrying out antigen retrieval, and carrying out closed treatment; (3) putting a perforated plate (3) in a longitudinal groove (5) to be fixed, wherein one surface covered with a flexible waterproof lining puts down; (4) adding various antibody solutions to be detected to different through holes (7) for incubation; (5) carrying out color development on the tissue section to be detected, sealing the tissue section, and observing. The invention also discloses a mold for preparing a cell tissue block. The high-throughput immunohistochemical detection method can simultaneously carry out the detection on various antibodies on a same slide and has a good application prospect.

The present invention relates to an in-vitro immunoassay method for detecting the presence of liver cancer cells in a subject. In the method of the present invention, antigen retrieval treatment based on heat-induced epitope retrieval method and antigen retrieval treatment based on protease-induced epitope retrieval method can be combined in the detection of glypican 3 antigen expression in liver cancer tissues to thereby detect the difference in the expression level or expression pattern of the glypican 3 antigen by immunohistochemical staining method. This enables samples, which has been determined by the conventional HIER method as highly expressing glypican 3, to be graded according to the expression level of glypican 3.

The invention discloses an antigen pre-processing method for preparing an antibody, and relating to the technical field of immunology. Purified antigen protein is processed by adopting an antigen retrieval solution in the pre-processing method. Aiming at the characteristics of an immunohistochemistry technology and disadvantages in the prior art, the invention aims to provide the antigen pre-processing method; and the antigen pre-processing method has the advantages of being simple and convenient to operate, low in production cost and the like. The key step of immunohistochemistry in the invention is antigen retrieval; the antigen is processed by selecting a corresponding antigen retrieval solution; the antigen pre-processing method, which can be applied to immunohistochemistry, is designed in a targeted manner, i.e., the immunohistochemistry retrieval manner of the antibody is selected according to the processing manner of the antigen; the antigen pre-processing method is simple and convenient to operate and low in production cost; and a novel effective way is provided for preparing the antibody applied to immunohistochemistry.

A method is disclosed for reversing fixation-induced cross-linking in tissue specimens that have been preserved for histological examination. The method involves placing the fixed tissue in a liquid under elevated temperature and pressure conditions that are sufficient to reverse the fixation-induced cross-linking, restore antigenicity to proteins, and permit improved molecular and proteomic analysis of the preserved tissue specimen. Methods are also disclosed for processing tissues for histological examination under elevated pressure conditions that enhance the perfusion of liquid reagents into the tissue and reduce overall processing times.

The present invention relates to the field of cancer. More specifically, the present invention provides methods and compositions useful for diagnosing or predicting cancer in a patient. In one embodiment, a method for identifying a patient as having cancer comprises the steps of (a) providing a formalin-fixed, paraffin-embedded or fresh frozen sample of patient tissue; (b) steaming the sample in antigen retrieval buffer; (c) incubating the sample in hydrochloric acid (HCl); (d) incubating the sample with an affinity reagent specific for 5hmC under conditions to form a complex between the affinity reagent and 5-hydroxymethylcytosine (5hmC) present in the sample; (e) detecting the complexes formed between 5hmC and the affinity reagent with secondary detection reagents; (f) quantifying 5hmC levels; and (g) identifying the patient as having cancer if the 5hmC levels in the sample are reduced as compared to a control.

The invention relates to a method for inactivating virus in vaccine production, belonging to the field of preparation of biological products. According to the method, the conditional parameters of temperature control, stirring rate, the flow rate of adding formaldehyde solution and the like during the inactivation process of the vaccine production can be improved according to the factors influencing the antigen stability, immunogenicity and antigen recovery rate during the inactivation of vaccine virus liquid are improved to acquire optimal inactivation conditions, the antigen loss during inactivation can be reduced, the antigen structure can be stabilized and the immunogenicity can be improved. According to the method, the quality stability and uniformity of products can be guaranteed, and the production cost can be lowered; the inactivation time can be shortened within 2h from 2.5h, the fatigue of people can be reduced, the production efficiency is improved, the intermediate product during vaccine production is prevented from being stored at room temperature for long time, and the quality of vaccines can be improved indirectly.

A method is disclosed for reversing fixation-induced cross-linking in tissue specimens that have been preserved for histological examination. The method involves placing the fixed tissue in a liquid under elevated temperature and pressure conditions that are sufficient to reverse the fixation-induced cross-linking, restore antigenicity to proteins, and permit improved molecular and proteomic analysis of the preserved tissue specimen. Methods are also disclosed for processing tissues for histological examination under elevated pressure conditions that enhance the perfusion of liquid reagents into the tissue and reduce overall processing times.

The present invention relates to an in-vitro immunoassay method for detecting the presence of liver cancer cells in a subject. In the method of the present invention, antigen retrieval treatment based on heat-induced epitope retrieval method and antigen retrieval treatment based on protease-induced epitope retrieval method can be combined in the detection of glypican 3 antigen expression in liver cancer tissues to thereby detect the difference in the expression level or expression pattern of the glypican 3 antigen by immunohistochemical staining method. This enables samples, which has been determined by the conventional HIER method as highly expressing glypican 3, to be graded according to the expression level of glypican 3.

The present invention relates to a method for enhancing immunoreactivity of a tissue or cell sample fixed in a fixing medium, a target retrieval composition and its use. The method comprises providing a carrier in a horizontal position, said carrier having thereon a tissue or cell sample, said tissue or cell sample being on top of the carrier; contacting substantially the tissue or cell sample side of the carrier with a buffered target retrieval solution, wherein the target retrieval solution remains otherwise exposed to the environment; heating the tissue or cell sample and the target retrieval solution to a temperature above 100° C. The invention furthermore concerns the automated immunohistochemical staining of said samples, in particular the reactivation of antigens masked by fixation.

The invention discloses a method for simultaneously labelling collagen type IV, macrophage and neovascularisation of tumors. The method comprises the following steps: 1. immobilizing tissue sections on a plus microscope slide processed by polylysine; 2. dewaxing the tissue sections in dimethylbenzene; 3. carrying out antigen retrieval; 4. washing the sections; 5. carrying out confining; 6. addingprimary antibodies; 7. washing the sections; 8. carrying out confining in the same way as the step 5; 9. using the fragment antigen-binding F(ab')2-QDs525 of goat anti-mouse immunoglobulin G labelledby quantum dot QDs525, the fragment antigen-binding F(ab')2-QDs585 of goat anti-rabbit immunoglobulin G labelled by quantum dot QDs585 and the fragment antigen-binding F(ab')2-QDs655 of rabbit anti-goat immunoglobulin G labelled by quantum dot QDs655 as secondary antibodies and dropwise adding the mixture after removing the confining liquid; 10. washing the sections; and 11. sealing the sections:preparing buffered glycerol with glycerol and 10ml of tris buffered saline (TBS) and storing the sections after sealing the sections with the buffered glycerol. The method is used for efficiently, accurately, rapidly and simultaneously detecting various components in tumor tissue microenvironment.

The invention relates to a suspended brain slice antigen retrieval pressing clamp which belongs to the field of medical experiment devices. The clamp consists of a rectangular frame (3) and a rectangular base frame (4), wherein the frame (3) is tightly compressed into a base groove (6) in match with the frame (3) in the base frame (4) through adjusting screw nuts (5); guide grooves (7) are formed in the four edges of the frame (3) and the four edges of the base frame (4); the guide grooves (7) in the frame (3) are matched with the guide grooves (7) in the base frame (4). By adopting the clamp, the defect that a suspended brain slice is easy to wrinkle in the conventional thermal retrieval process is overcome, the suspended brain slice antigen retrieval pressing clamp provided by the invention is simple in structure and convenient to operate, the antigen retrieval of the 'suspended bran slice' can be achieved, and a brain tissue section is prevented from wrinkling.

The invention relates to the technical field of immunohistochemistry and discloses EDTA antigen retrieval liquid. Each 1,000 mL of EDTA antigen retrieval liquid is prepared from 1.20-1.25 g of Tris-Base, 0.35-0.40 g of EDTA, 400-600 microliters of twain-20, 50-500 mL of triethanolamine or ethanediol or triethanolamine and the balance single steaming water. The EDTA antigen retrieval liquid can achieve infiltration, is good in retrieval effect and can retrieve tissue slices with a little amount, the retrieved tissue slices can be dyed easily, and the dyeing effect is good. The retrieval liquid is not prone to volatilization and can be used in cooperation with a full-automatic immunohistochemical instrument.

Heat induced antigen retrieval systems for biological specimens may include a sealable heating pressure chamber, a programmable process controller, a nonpareil operating element, and perhaps even a substantially user-disencumbering autonomous processing component of a plurality of biological samples perhaps using various user selected protocols, and the like.

Solutions exhibiting little or no evaporative loss at elevated temperatures, i.e., in excess of 100° C., are employed in place of conventional aqueous-based antigen retrieval solutions.

The invention belongs to the technical field of immunohistochemical staining. An instant preparation and instant use module is arranged on an immunohistochemical staining machine to achieve the instant preparation and instant use of a reagent required in the staining process of a tissue slide. After the tissue slide is stained, a cooling structure is arranged under a staining working platform andused for cooling the staining working platform. Meanwhile, an antigen retrieval module is arranged in the machine and can conduct high-temperature water bath antigen retrieval on a tissue slice.

Disclosed are a pretreatment solution for immunohistochemical staining, which elutes a paraffin-containing embedding medium from a glass slide with a tissue specimen embedded in the medium, and retrieves antigenicity of the tissue specimen, and which is usable three or more times, and a pretreatment solution concentrate for immunohistochemical staining which allows ready preparation of the pretreatment solution. The pretreatment solution for immunohistochemical staining contains an antigen retrieving agent, particular nonionic surfactants, and cyclodextrin or a derivative thereof, with the balance being not less than 80 mass % of water. The content of the antigen retrieval agent is such that the pH of the pretreatment solution is in a predetermined range, and the content of cyclodextrin or a derivative thereof is a particular amount.

The invention discloses an immunofluorescent method for detecting plant gibberellin, and belongs to the field of plant biotechnology. The immunofluorescent method for detecting plant gibberellin comprises the following steps: S1, dewaxing rehydration; S2, antigen retrieval; S3, autofluorescence quenching; S4, serum sealing; S5, addition of one antibody; S6, addition of two antibodies; S7, DAPI cell nucleus redyeing; S8, mounting; and S9, microscopic examination. The method is simple, rapid, safe and non-toxic, the detection cost is reduced effectively, histocytes are positioned by using DAPI redyeing cell nucleuses, and the position of antigen distribution can be observed and determined preferably; the distribution condition of gibberellin inside plant tissues can be observed through a fluorescence microscope after dyeing; the original states of plant tissues and cells are maintained to the maximum extent in the experimental process; and the biological activity and immunocompetence canbe maintained after marking.

The invention discloses a method for observing a microtubule structure of a cytoskeleton in a liver tissue of animals. The method comprises the following steps: (1) cleaning the liver tissue of animals with PBS, fixing for 8-12h with 4v / v% of paraformaldehyde water solution, and carrying out paraffin embedding and serial sectioning; (2) dewaxing the obtained paraffin sections, and carrying out antigen retrieval, closing and fluorescently labeled antibody incubation; (3) for specimens sealed by glycerol, observing through a laser scanning confocal microscope within 24h, respectively exciting 488 and 4' 6-diamidino-2-phenylindole under 488nm and 405nm, and observing and shooting at once at room temperature in a dark room environment. The method is fast, simple, intuitive and accurate, is strong in practicality and extensibility, and can research the dose-effect changing relationship of the cytoskeleton under stress of pollutants.

The invention relates to the field of molecular biology, and provides a RNA-protein-DNA in-situ multiple staining method of a tissue chip. The method comprises the following steps: subjecting a tissue chip to RNA in-situ hybridization, immumohistochemical staining, and DNA in-situ hybridization in sequence; during the processes of RNA in-situ hybridization, immumohistochemical staining, and DNA in-situ hybridization, subjecting the tissue chip to molecule crosslinking melting or antigen retrieval by adopting an epitope antigen retrieving steamer; and during the processes of RNA in-situ hybridization and immumohistochemical staining, sealing the chip by gelatin glycerol after color development. The provided method can detect one tissue chip in three molecular levels (RNA, protein, and DNA), can obtain more comprehensive biological information, and reduces the cost of obtaining the biological information.

Compositions and methods for preparing a sample for immunological staining are provided. Compositions include kits comprising a first solution comprising a surfactant and a second solution comprising a chaotropic agent. Methods comprise contacting a sample, such as cells or tissues, with a first solution comprising a surfactant and then contacting the sample with a second solution comprising a chaotropic agent. The method does not require extreme heat for antigen retrieval and therefore, maintains the cellular morphology of the sample.

The invention discloses a concentration method of Seneca virus. The method comprises pretreatment of a Seneca virus liquid, and concentration after the pretreatment, wherein the concentration sequentially comprises to filtration concentration, filter wash and filtration concentration. The invention relates to an SVV antigen concentration and purification method suitable for large-scale industrialproduction. The method subjects an SVV antigen to concentration and purification by combining tangential flow filtration and filter wash, and has the characteristics of convenient operation, easy amplification, mild and stable process, high antigen recovery rate and the like. The method can be used for concentration and purification of live viruses and inactivated viruses, has an antigen recoveryrate of greater than 80% and a protein removal rate of greater than 90%, and can prepare a purified SVV antigen or provide raw materials for subsequent further fine purification of the antigen.

The invention discloses an automatic dewaxing antigen retrieval system, which comprises a reaction device, a dewaxing reagent injection device, a first reagent heating device, an eluent injection device, an enzyme diluent injection device, a concentrated enzyme reagent injection device, a second reagent heating device, a permeation agent injection device, a third reagent heating device, a gas injection device, an anhydrous ethanol injection device, a deionized water injection device and a waste liquid collection device, wherein the reaction device comprises a reaction box, a fan and a reaction heater, and various injection devices are respectively connected to the reaction box and supply various reagents and the reaction pressure required by the dewaxing reaction in the reaction box. According to the present invention, the automatic dewaxing antigen retrieval system has advantages of simple structure, convenient operation, stable performance and low dewaxing cost, and can rapidly perform dewaxing treatment on tissue slices.