29 results about "Paraffin embedded" patented technology

The invention discloses application of jellyfish and snow rabbits in inhibition of ERK phosphorylation level and MAPK pathway and an evaluation method of the jellyfish and snow rabbits, and relates to the technical field of biological medicines.The method comprises the following steps that 1, a treated jellyfish and snow rabbits breast cancer cell group and a control breast cancer cell group are established; and step 2, respectively dividing the two groups of samples in the step 1 into two parts, fixing one part in paraformaldehyde with the mass concentration of 10%, treating, embedding in paraffin for immunohistochemical analysis, and quickly freezing the other part for transcriptomics analysis and the like. According to the invention, UPLC-MS is adopted to determine the main components of SMM, the influence of SMM on breast cancer (BC) is evaluated through in-vivo experiments, in order to deeply clarify the mechanism, network pharmacology and transcriptomics analysis are integrated, finally, immunohistochemistry (IHC) is adopted to verify and explore the exact breast cancer resisting mechanism of SMM, and a scientific basis is provided for potential clinical application of SMM.

The present disclosure encompasses methods for molecularly subtyping formalin-fixed paraffin-embedded tumor samples. The disclosure works particularly well for old and degraded (archival) samples for which standard methods are unfeasible. Further, the methods disclosed allow for the correlation of patient outcome data with the molecular subtype of the tumor and provides a wealth of information which will guide treatment decisions and / or selection of therapeutic agents.

The disclosure provides methods for the use of gene expression measurements to classify or identify neuroendocrine cancer in samples obtained from a subject in a clinical setting, such as in cases of formalin fixed, paraffin embedded (FFPE) samples.

Aspects of the present disclosure are directed to methods, compositions, and kits for detection and analysis of DNA and RNA cytosine methylation and / or RNA pseudouridylation. Certain aspects include methods, compositions and kits useful in bisulfite sequencing of methylated and / or pseudouridinylated nucleic acids, including methylated and / or pseudouridinylated nucleic acids from low-input samples such as cell-free DNA (cfDNA) and cell-free RNA, long fragment polynucleotides, and / or cfDNA and DNA from formalin-fixed paraffin-embedded (FFPE) samples. Also disclosed herein are methods, compositions and kits useful in bisulfite sequencing of methylated and / or pseudouridinylated nucleic acids with low nucleic acid damage, higher true positive results, and / or lower false negative results.

This invention discloses a bone marrow cell paraffin block, its preparation method, and its application, belonging to the field of pathological detection technology. The preparation method includes the following steps: Red blood cell lysis: Bone marrow aspirate is taken, and a precipitate containing cells and sediment is separated. A red blood cell lysis agent is added to lyse the red blood cells, and a lysed precipitate containing unlysed cells and sediment is separated. Formation: The lysed precipitate is resuspended in formalin solution for fixation, the supernatant is removed, and the precipitate is resuspended in agar solution. The agar is cooled and solidified, and then fixed again in neutral formalin. Dehydration: The solidified agar block is dehydrated. Embedding: The dehydrated agar block is embedded in paraffin to form a paraffin block. The cell paraffin block prepared by this method can supplement the diagnosis when bone marrow smears or bone marrow biopsy samples are not ideally prepared.

Aspects of the present disclosure are directed to methods, compositions, and kits for detection and analysis of DNA and RNA cytosine methylation and / or RNA pseudouridylation. Certain aspects include methods, compositions and kits useful in bisulfite sequencing of methylated and / or pseudouridinylated nucleic acids, including methylated and / or pseudouridinylated nucleic acids from low-input samples such as cell-free DNA (cfDNA) and cell-free RNA, long fragment polynucleotides, and / or cfDNA and DNA from formalin-fixed paraffin-embedded (FFPE) samples. Also disclosed herein are methods, compositions and kits useful in bisulfite sequencing of methylated and / or pseudouridinylated nucleic acids with low nucleic acid damage, higher true positive results, and / or lower false negative results.

The application discloses a fine segmentation method for prostate and internal lesion areas based on large pathological sections, and specific steps are as follows: step 1, sequentially performing fixation, paraffin embedding, continuous transverse sectioning and HE staining operations on the whole tissue; step 2, extracting an HE staining image; step 3, sequentially scanning the pathological sections in step 2 into digital pathology; step 4, processing the digital pathology; step 5, performing image registration and three-dimensional image reconstruction on analysis results of multiple pathological sections processed in step 4; step 6, training a magnetic resonance multi-modal sequence segmentation model; step 7, extracting features of normal prostate tissue and lesion tissue in three sequences; step 8, acquiring high-risk factor information and quantifying features; step 9, constructing a sample feature matrix; and step 10, predicting the malignancy degree of prostate cancer. The method can realize more accurate benign and malignant evaluation of prostate lesions and prediction of the malignancy degree of prostate cancer.

The invention relates to the technical field of animal histological analysis technology and skin appendage application, in particular to a goose skin hair follicle microsection manufacturing and evaluating method, which specifically comprises the following steps: selecting back skin of a Sanhua goose, avoiding dehairing injury, cutting a 5 * 5mm full-thickness skin sample by using a scalpel, and taking out the full-thickness skin sample; a sample is immediately put into 4% paraformaldehyde to be fixed for 24 hours, stepped ethanol dehydration is adopted, tissue brittleness is reduced, paraffin embedding optimization is adopted, so that a hair follicle structure slice is clear in layer, the method is easy and convenient to operate, tissue treatment procedures do not need to be optimized, it is guaranteed that the thickness of the hair follicle slice is more uniform, and the quality of the hair follicle slice is improved. The number of primary hair follicles and hair follicles is uniform, recording is accurate, hair follicle slice observation and number analysis are facilitated, and the method plays an important role in goose breeding and down feather quality evaluation.

The application relates to an experimental method for improving the accuracy of laboratory pollution monitoring and the quality control capability in molecular pathology detection. The method comprises the following steps: preparing a blank wax block without any tissue component while preparing a tissue paraffin-embedded sample to be detected; slicing the tissue paraffin-embedded sample and the blank wax block respectively; extracting nucleic acid of the tissue sample and nucleic acid of the blank wax block respectively; carrying out nucleic acid amplification detection by taking the nucleic acid of the tissue sample as a sample to be detected, taking the nucleic acid of the blank wax block as a negative whole-process control, and combining a positive control; judging the pollution and analyzing the quality control of the whole experimental process according to the detection result of the negative whole-process control of the blank wax block, so that the whole-process pollution risk of molecular pathology detection is continuously monitored. The application has the effects of improving the accuracy of laboratory pollution monitoring, the reliability of experimental results and the whole-process quality control capability of the laboratory.

The invention discloses application of a detection reagent (a reagent for simultaneously detecting MET, PCDHGA3 and FAM3C) in detection of ZM fusion gene (PTPRZ1-MET fusion gene) positive secondary glioblastoma (sGBM), belongs to the technical field of biological medicine, and aims to solve the problems that an existing ZM fusion detection method is complex in operation, poor in specificity, not suitable for conventional clinical detection and the like. Based on high-throughput RNA sequencing and machine learning algorithm (XGBoost) analysis, it is found for the first time that a combined expression profile of the three genes MET, PCDHGA3 and FAM3C can remarkably distinguish ZM fusion positive and negative samples, the diagnosis accuracy (AUC) of the combined expression profile reaches up to 99.8%, and the combined expression profile has high sensitivity and specificity. The detection product is suitable for paraffin embedding (FFPE) pathological tissue samples and frozen samples, detection can be completed in combination with mature platforms such as multiple immunohistochemistry or high-throughput RNA sequencing, and good sample compatibility and operation feasibility are achieved.

The application discloses a method for relieving obesity cardiomyopathy lipid toxicity by Chemerin and belongs to the technical field of pharmacological research. Mice are selected to establish an obesity mouse model, and are equally divided into a control group, a high-fat diet group, an AAV CMKLR1 virus group, and a high-fat + AAV CMKLR1 virus group. In succession, mouse heart ultrasound, TSE system detection, glucose tolerance test, and insulin tolerance test are carried out, and mouse blood glucose is measured at intervals. Then, tissue protein is extracted for protein immunoblotting, and tissue RNA is extracted for reverse transcription, and then qPCR is used to detect the expression of related genes. After paraffin embedding and slicing of mouse tissues, HE, WGA, immunofluorescence and other pathological staining are carried out. MTBE method is used for lipid extraction and detection of lipid content. ELISA method is used to measure serum Chemerin, and Acyl-RAC method is used to detect protein palmitoylation. Finally, the experimental results are analyzed to draw a conclusion. The present application proves by clinical data and mouse models that Chemerin compensatorily increases in the process of obesity, and Chemerin can relieve cardiac lipid toxicity caused by obesity and protect cardiac function.

The present application belongs to the technical field of quality control materials. Disclosed are a quality control material freeze-dried powder as well as a preparation method therefor and the use thereof. The quality control material freeze-dried powder is obtained by freeze-drying cell pellets containing cell fragments, the cell pellets being obtained by sectioning, deparaffinizing and hydrating a paraffin-embedded cell sample and then centrifuging same. The quality control material freeze-dried powder prepared by using the preparation method in the present application can reduce the loss of cell fragments during the freeze-drying process, so as to ensure the high recovery rate of cells, can avoid performance change or protein loss of cell fragments during the freeze-drying process, and can ensure that the immunohistochemical staining performance of reconstituted cell pellets after freeze-drying remains unchanged. The quality control material freeze-dried powder obtained by using the preparation method in the present application has extended storage time and facilitates transport, and the reconstituted cell suspension has good dispersibility, retains key target substances, and exhibits normal immunohistochemical staining performance.

The invention relates to the technical field of organoid embedding, in particular to a mold device for organoid embedding, and solves the problems that in the prior art, the whole dehydration-embedding process cannot be simplified, and the risk of organoid damage and orientation offset exists. The mold device comprises a bottom plate and a top cover, and the bottom plate and the top cover are buckled in a detachable mode and form an embedding groove; a plurality of dehydration pore channels are formed in the mold device, so that the embedding groove is communicated with the outside; the top cover is provided with a wax liquid guide mechanism, and the demolding mechanism comprises a knob, a threaded rod and a demolding unit; the knob is rotationally connected with the bottom face of the bottom plate, the axis of the top face of the knob is fixedly connected with one end of the threaded rod, the threaded rod is sleeved with the demolding unit, and the threaded rod rotates to enable the demolding unit to ascend and descend. According to the device disclosed by the invention, by integrating dual functions of dehydration treatment and paraffin embedding, full-process in-situ operation from dehydration to forming of the organ-like sample is realized, a manual transfer link is thoroughly eliminated, and structural damage and orientation deviation of the pre-embedded sample are avoided.

The invention discloses a large tissue slice embedding and clamping assembly, and relates to the technical field of embedding boxes, the large tissue slice embedding and clamping assembly comprises a clamping adaptive seat and an embedding sheet, two parallel clamping surfaces are arranged on two sides of the clamping adaptive seat, the clamping surfaces are used for clamping a sample clamp of an automatic pathological paraffin embedding wheel type slicer, and the embedding sheet is arranged on the clamping adaptive seat. The tops of the two parallel clamping faces are connected through a positioning face, the end faces of the bottoms of the two parallel clamping faces are flush and can abut against the bottom face of the sample clamp, an embedding piece is detachably connected to the positioning face, and a large tissue slice wax block is arranged on the embedding piece. The clamping adaptive seat is fixedly connected with a sample clamp of a slicing machine, and a plurality of embedding pieces can be replaced for slicing through one-time clamping of the clamping adaptive seat, so that a large tissue slice wax block can be quickly clamped, the position of the rear embedding piece can be relatively fixed, the section angle can be kept on the same section after repeated slicing, and the slicing efficiency is greatly improved. And the phenomenon that the focus tissue is cut by repeatedly adjusting the angle of the tangent plane is avoided.

The present invention pertains to a method of preparing a frozen biological sample, comprising the steps of fixing the biological sample with a non-crosslinking fixative solution, incubating the fixed biological sample in an aqueous solution comprising a cryoprotectant, and freezing the cryoprotected biological sample. The method advantageously allows to preserve both, morphology of the sample as well as biological components such as nucleic acids and proteins, in high quality for subsequent analysis. The method is robust, simple and neither requires laborious steps associated with paraffin-embedding nor immediate freezing of the sample. Also provided are advantageous uses and kits.

Methods and reagents suitable for in vitro diagnostic assay comprising a qualitative immunohistochemical assay using anti-Globo H antibodies and / or binding fragments thereof are provided. The method comprises the detection of Globo-H expression levels in formalin-fixed, paraffin-embedded (FFPE) cancer tissue using a visualization system. The Globo-H expression can be determined by using tumor scoring showing partial or complete staining at any intensity.

The invention provides a preparation method of a paraffin-embedded section of decalcified bone tissue, which belongs to the field of paraffin section manufacturing and comprises the steps of fixation, decalcification, gradient concentration ethanol treatment, embedding, freezing, slicing and the like. The method for treating the glass slide is simple and low in cost, a uniform film can be formed, the section keeps good adhesiveness in the embedding and dyeing process, meanwhile, the solvent resistance is good, the problem of section falling caused by xylene dewaxing is solved, tissue loss can be remarkably reduced, and the experiment stability and repeatability are improved. In addition, a temperature gradient wax dipping method is also used, so that the permeation rate of paraffin inside and on the surface of the tissue is more uniform, a uniform and stable paraffin embedding matrix is finally formed, the slice quality is improved, and the dyeing and microscopic observation effects are clearer.

The present disclosure discloses a library construction method for detecting endometrial cancer-related gene mutations based on high-throughput sequencing, and belongs to the field of biotechnology. The method can detect the mutation types of endometrial cancer-related genes MSH2, PMS2, MLH1, MSH6 EPCAM, TP53, POLE, and PTEN in surgically removed fresh pathological tissues, formaldehyde-fixed and paraffin-embedded pathological tissues, paraffin sections, and specimens of whole blood, plasma, serum, and pleural effusion, etc. It may be used for multiple target sequences in a single tube to quickly complete the library construction. The entire library construction process only takes 3 hours, and the manual operation only needs 30 minutes. Combined with high-throughput sequencing, the platform may effectively solve the current difficulty in the detection of somatic multi-gene all-exon mutations in clinical endometrial cancer samples based on small numbers of clinical samples, and the cost is low.

The present disclosure is directed to a non-enzymatic dissociation method which facilitates the dissociation of one or more formalin-fixed paraffin-embedded tissue samples into dissociated single cells. The present disclosure is also directed to methods of single cell analysis, i.e., methods of analyzing and / or measuring target components (e.g., biomolecules such as, but not limited to, polypeptides, polynucleotides, small molecules, and the like) on or in cells non-enzymatically dissociated from one or more FFPE tissue samples.

The invention belongs to the technical field of animal tissue sections, and particularly relates to an optic nerve paraffin section, a preparation method and a kit, the preparation method comprises the following steps: S1, obtaining an eyeball, and placing the eyeball in a stationary liquid for preliminary fixation; s2, finding the optic nerve, and locally marking the proximal end of the optic nerve by using a hematoxylin solution; s3, carrying out gradient dehydration treatment, transparency treatment and wax dipping treatment on the marked optic nerve tissue blocks; s4, carrying out paraffin embedding on the tissue block subjected to paraffin dipping; and S5, slicing the embedded block, and carrying out HE dyeing to obtain the paraffin section. According to the preparation method, the preparation method of the paraffin section is improved, so that the visibility, the positioning accuracy and the structural integrity of optic nerves are remarkably improved, and the optic nerve paraffin section with better integrity and definition is obtained.

The embodiment of the utility model discloses an embedding box for fixing pathological tissues, the embedding box is a box body, the box body comprises a box cover and a containing box detachably connected with the box cover, a plurality of partition plates are arranged in the containing box, and a plurality of containing grooves are formed; a plurality of through holes are formed in the bottoms of the box cover and the accommodating box; a pathological tissue fixing piece is arranged on the inner side of the box cover and corresponds to the upper part of the accommodating groove, and the pathological tissue fixing piece is matched with the bottom of the accommodating groove and can fix the pathological tissue in the accommodating groove. According to the embedding box for fixing the pathological tissue, the pathological tissue fixing pieces are correspondingly arranged in the containing groove, so that the direction of the pathological tissue of the pathological tissue is convenient to fix, pathological tissue materials are convenient to fully soak, and the position of the pathological tissue is convenient to identify; the diseased tissue part is easy to identify, the target diseased tissue is convenient to cut after paraffin embedding, and a good foundation is laid for accurate detection.

This invention discloses a method for extracting cell nuclei from formaldehyde-fixed and paraffin-embedded samples, comprising: (1) extracting cell nuclei: cutting PA / PFA-fixed or FFPE samples into pieces, adding cell nuclei extraction solution, incubating, collecting the supernatant, centrifuging, and discarding the supernatant; (2) washing cell nuclei: washing the cell nuclei precipitate from step 1) with PBS mix, centrifuging, discarding the supernatant, and resuspending the cell nuclei with PBS mix. The cell nuclei extracted by this method have advantages such as good morphological integrity, high purity, good RNA integrity, and high RNA yield, which are of great significance for transcriptomics studies of PA / PFA-fixed and FFPE samples.

Provided herein are methods for detecting the presence or absence of a chromosome rearrangement in a formalin-fixed paraffin embedded (FFPE) sample with no detectable genomic variant associated with cancer. In certain embodiments, the methods include selecting a sample and performing a nucleic acid analysis on the selected sample and detecting whether a chromosome rearrangement is present or absent in the selected sample according to the nucleic acid analysis. In preferred embodiments, the nucleic acid analysis is a method that preserves spatial-proximal contiguity information.

A method for processing tissue section images is provided. The method includes: (S1) providing a paraffin-embedded tissue block; (S2) determining at least three first coordinates on tissue sections to be sliced off from the paraffin-embedded tissue block, ablating at the first coordinates to form marker points on the tissue sections, and slicing the ablated tissue sections off from the paraffin-embedded tissue block; (S3) acquiring a slide image of each of the ablated tissue sections; and (S4) determining a second coordinate corresponding to each marker point on the slide image, comparing for each marker point the second coordinate with the first coordinate, and processing the slide image according to the comparison results. A system for performing the method for processing tissue section images is also provided.

The application relates to the field of organoid embedding technology, and particularly relates to a mold device for organoid embedding, which solves the problems of the inability to simplify the whole process of 'dehydration-embedding', the risk of organoid damage and position deviation in the prior art, and comprises a bottom plate and a top cover, the bottom plate and the top cover are detachably buckled and form an embedding groove; a plurality of dehydration channels are formed in the mold device to enable the embedding groove to communicate with the outside; the top cover is provided with a wax liquid flow guide mechanism, and the mold device further comprises a demolding mechanism, the demolding mechanism comprises a knob, a threaded rod and a demolding unit; the knob is rotationally connected to the bottom surface of the bottom plate, the top surface of the knob is fixedly connected with one end of the threaded rod, the demolding unit is sleeved on the threaded rod, and the threaded rod is rotated to lift or lower the demolding unit. The device has the dual functions of dehydration treatment and paraffin embedding, realizes in-situ operation of the whole process from dehydration to molding of the organoid sample, completely eliminates the manual transfer link, and avoids structural damage and position deviation of the pre-embedded sample.

The application discloses a kind of construction methods of sunlight dermatitis mouse model, specifically belongs to animal experiment model experimental technical field, the construction method includes the following steps: S01, ultraviolet lamp intensity measurement and selection;S02, the establishment of sunlight dermatitis model;S03, fresh tissue paraffin embedding section;S04, wax tissue section respectively by H-E staining experiment, Masson staining experiment, immunohistochemical staining experiment, transmission electron microscopy sheet observation, obtain the structure characteristic change of material organization.This construction method is by controlling the consistency of keeping radiation dose, to restore actual situation to a greater extent, provides real and reliable basis for developing sunlight dermatitis drug.

Provided herein is a multiplexed immunohistochemistry staining and digital pathology analysis dual-process quality control product, comprising a formalin-fixed paraffin-embedded (FFPE) tonsil sample, and a cell dispersion zone located in the center of the FFPE tonsil sample, the cell dispersion zone comprising dispersed PBMCs embedded with agarose gel and embedded with paraffin. Also provided herein are methods of preparing and using the quality control product.

The present invention relates to the technical field of dewaxing buffers, specifically disclosing a water-soluble immunohistochemical dewaxing buffer and its application. The water-soluble immunohistochemical dewaxing buffer comprises the following components by weight: 1 to 4 parts of a surfactant; 0.5 to 6 parts of a water-soluble organic solvent composition; and 80 to 100 parts of water. The water-soluble immunohistochemical dewaxing buffer can dissolve the paraffin embedded in tissue sections at room temperature within 15 to 30 minutes, and the dewaxed tissue can be directly subjected to immunohistochemical staining without requiring hydration.