117 results about "Tissue Chip" patented technology

The invention discloses a CRISPR-Cas9 system for knocking out the gene GRIN2D and its application. The CRISPR-Cas9 system comprises Cas9 and gRNA specifically targeting to the gene GRIN2D. The invention also discloses application of the CRISPR-Cas9 system to preparation of drugs used for treating cancer and application of the gene GRIN2D to diagnostic kits and medicines used for precise treatmentof stomach cancer and other tumors. In virtue of a clinical sample tissue chip of stomach cancer, the gene GRIN2D is proved to be increased in expression in stomach cancer tissue and be related to prognosis according to immunohistochemical results. The CRISPR-Cas9 system provided by the invention can highly efficiently knock out the highly-expressed gene GRIN2D in stomach cancer and inhibit the proliferation and migration / invasion of gastric cancer cells, and is simple to operate and high in knockout efficiency. The CRISPR-Cas9 system is expected to be applied to diagnosis and treatment of stomach cancer and other tumors with over-expressed GRIN2D. The CRISPR-Cas9 system is applicable to a plurality of tumors with abnormal expression of GRIN2D.

Three kinds of tissues including cancer tissue, precancerosis and corresponding normal tissue are sliced up, dyed, marked, and positioned. Receptor holes are prepared by leading designed lattice array mould paper to paste on surface of wax block of receptor. Wax block with tissue core bar is prepared by using perforating needle and puncture needle for tissue. Common cancer such as lung cancer, nasopharyngeal carcinoma, oesophagus cancer etc. and having integrated clinical data and pathology features are selected. Through in situ hybridization, testing mRNA of relevant gene and expression of protein on tissue chip, consistent result between the invented product and traditional test is validated. In the product, cellular morphology is clear and even, and there is no fallen off tissue point. The invention is applicable to filter cancers, early diagnosis and forecasting prognosis.

A tissue chip for detecting autoantibody is composed of substrate and the matrix on the substrate. Said matrix may be paraffin wax coated tissue, frozen fresh tissue, or cell specimen. An indirect immunofluorescence technique is used to detect, screen and analyze a great deal of autoantibodies in the blood serum of autoimmunopathy patient.

The invention discloses a preparing method for an olefin tissue core which includes the steps of customizing an olefin model, customizing a tissue array hole plate, customizing a mandrel, customizing a sampling needle, customizing the olefin block of the tissue chip array of a receptor, sampling the olefin block of a supplier, preparing the olefin block of the tissue chip of the supplier, etc. The preparing method is simple and easy with good experimental result.

The invention provides a method for embedding wax tissue chip, which is characterized in that the method comprises the detailed steps of cutting small tissue blocks on the wax tissue, storing the small tissue blocks in a tissue chip storage plate and numbering the small tissue blocks; arranging the tissue chip storage plate on a burning platform till the wax composition is melted in the small tissue blocks; taking the small tissue blocks out of the tissue chip storage plate and arranging the small tissue blocks in a tissue chip embedding plate; arranging the tissue chip embedding plate in a rectangular wax mold assembled by two vertically bent copper bars; taking the wax out and arranging the wax into a wax kettle; heating and melting the wax by an electric furnace; adding the melted wax into the wax mold along one end of the wax mold till the wax mold is fully filled; removing the wax mold, separating the tissue chip embedding plate from the wax; and slicing along the direction in parallel to the even surface of the tissue chip. The method has the advantages of being low in cost, conveniently, quickly and freely collecting the chip sample, freely storing the samples in the storage plate, and having repeatable usage.

The invention relates to the biochips field, in particular to a tissue chip used for functional genome research and the preparation method as well as the application thereof. The tissue chip of the invention includes a substrate and a paraffin section arranged on the surface of the substrate, wherein, the paraffin section is provided with tissue sample points which are in different development stages and array according to the tissue dynamic development process. Because the tissue chip of the invention can be combined with a gene chip technology, the space-time expression of a difference expressive gene screened out by the chip technology in the tissue dynamic development process is validated and researched in the high-quality, economic and available way.

The invention which belongs to the biotechnological field concretely relates to a method for screening a tumor specificity target and a targeting ligand with a tissue chip. The invention which aims to overcome disadvantages of low medicine or targeting treatment versatility, large gap between a screening system and in-vivo living environment of a tumor, small screening throughout, and time and labor consumption existing in the prior art provides a method for screening the tumor specificity target and the targeting ligand with the tissue chip. The targeting ligand screened in the invention has the advantages of high specificity, realization of tumor cell penetrating, no immunization reaction causing, and high versatility. The screening method maintains the original state of the in-vivo grown tumor; and compared with tumor-bearing mouse screening systems, the screening method has the advantages of animal breeding step omitting, time, labor saving and cost saving, and strong experiment result comparability. Compared with traditional screening systems, the screening method of the invention allows the tumor specificity target analysis and the targeting research to be simultaneously carried out, and the verification process is simple.

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 provides application of SMOC2 gene (SPARC related modular calcium binding2, with the Chinese name of secreting modular calcium binding protein 2; smooth muscle related protein 2) in preparation of medicine for detecting or treating endometrial cancer and ovarian cancer. A tissue chip comprising 157 endometrial cancer tissue specimens, 30 normal proliferative phase endometrium tissue specimens and 30 normal secretory phase endometrium tissue specimens is established by utilizing a tissue microarray technology, the expression conditions of SMOC2 in the specimens are detected by utilizing the immunohistochemical technique, the positive expression rate of the SMOC2 in the normal tissue of the endometrium is far lower than the positive expression rate in the endometrial cancer, and the expression intensity is positively correlated to muscular invasion depth of the endometrial cancer and tumor grading of the endometrial cancer. Therefore, the SMOC2 gene can be used for preparing the medicine for detecting or treating endometrial cancer. The medicine is reliable in experimental result and high in repeatability and has good clinical application prospect and good application values and social benefits.

The invention provides a simple inoculation method for a paraffin tissue chip. The method is characterized by comprising the steps of: conducting sampling on a pathologic tissue wax block; heating the wax to a melting state, placing a wax pattern preparation frame on a plastic wax template, pouring wax into the wax pattern preparation frame for polymerization and cooling, and removing the plastic wax template; putting the obtained wax pattern and the wax pattern preparation frame in an incubator, and taking the wax pattern out after it softens, carrying out drilling with the inoculation scope of the wax pattern, and planting a chip tissue into a hole corresponding to the wax pattern; placing the inoculation face of the obtained wax pattern on a flat ironing platform, after melting of the wax, stopping heating and leaving the wax to natural cooling; reheating the flat ironing platform, and removing the wax pattern and the wax pattern preparation frame; placing the wax pattern and the wax pattern preparation frame into a refrigerator, then taking them out and separating them, and slicing the obtained wax pattern so as to obtain a paraffin tissue chip. The method of the invention has the advantages of low cost, convenience, rapidity, high yield, and sampling diversity, and can be popularized to every pathology department as well as related research units.

The present invention provides a technology and a method for preparing a high-throughput exfoliated cell chip. According to the present invention, a cervix brush is adopted to collect cervix exfoliated cells of 26 cervical lesions patients and are immediately placed into a cell preservation solution, centrifugation is performed, the supernatant is removed, a cell immobilization solution is added to immobilize cell precipitate, dehydration transparency and dipping in wax are performed, a tissue chip instrument is adopted to prepare an exfoliated cell paraffin chip and an exfoliated cell paraffin block array, and paraffin slicing is performed to prepare the exfoliated cell chip; and the defect of the conventional cytology detection is overcome, and the prepared exfoliated cell chip can be used for pathological diagnosis, IHC, in situ hybridization and other molecular medicine researches.

This invention discloses one tumor transfer mimic organism chip, which is processed by the following steps: selecting the supply wax organism specimen through each phase in transfer process and through normal HE dyeing assistant immune set; positioning tumor transfer process each point for label; collecting labeled organism to form chip matrix slice; then processing the silicon slice through process. This invention can test tumor transfer process relative gene expression and its status and filter prediction factors and after factors to guide tumor personal medical and target medicine.

The invention discloses a kit for constructing forecasting NSCLC (non-small-cell lung cancer) patient prognosis and guiding clinic application of zoledronic acid and application of osteopontin (OPN) acting as a specific marker of M2 type macrophage; the application of the kit provided by the invention is evaluation of the NSCLC patient prognosis by using the expression of OPN in an immunofluorescence double standard technology detection tissue chip in TAMs and tumor cells, and comprehensive measures are taken in real time to interfere aiming at the detection result of the kit, so that the long-term survival rate of patients is increased, and the prognosis is improved; the method is simple and convenient and can be widely applied to clinic.

An instant powdered coriander microcapsule as flavouring is prepared from fresh coriander through prebreaking, pressing to obtain coarse juice, filter, superfine pulverizing to obtain its tissue chips, heating, filtering to obtain clear juice, mixing it with arabic gum and malt dextrin, and spray drying.

The invention discloses a tissue chip making system based on image recognition and positioning and a chip making method thereof. The chip making system comprises a cutting system and a computer control system; the cutting system comprises a numerical control cutting machine, an XYZ axis translation work table, a 360-degree-rotation rotary table, a receptor wax block frame, a freezing table and animage recognition and positioning module. The chip making process of the chip making system comprises sample placing, sample position identification, tissue sample image acquisition, tissue sample image processing, setting of cutting parameters, cutting of a tissue sample, tissue chip information identification and storage, and obtaining of the tissue chip with traceability of information. The method obtains a chip taking area by visual identification, has the advantages of high degree of automation and high work efficiency, through linear cutting of the tissue sample, tissue damage is effectively reduced, and the chip taking efficiency and success rate are improved; a database is established for management of the information of the tissue chip, so as to be easy to follow-up queries.

It is organization chip process method and its apparatus which comprises sample device 1 and process device 11 and in detail comprises control pressure bar1, guiding module 3, fixed screw 4, sample needle 5 of needle chip 11, reset pressure spring 6, position bolt 7, guiding bar 9, control bolt 12, needle chip control panel 14, array distribution board 15, L-shape board 16 and guiding bar 17. It is to centrally sample through sample device and then to distribute a group of sample needles 5 orderly on the array distribution board 15, and to inject olefin into all the organic samples through pushdown needle chip control board 14 and it is finished when the olefin is solidified. This apparatus in this invention is in no need of olefin block process and stiletto procedure and accuracy position system.

The invention provides a technology and method for preparing high throughput needle biopsy tissue chip. The method comprises the following steps: collecting 42 residual paraffin-embedded tissue samples of needle biopsy of patients who have been diagnosed as the breast cancer patients through pathological examination on mammary gland needle biopsy tissues; taking the corresponding normal pathological HE stained sections as the guide, cutting the paraffin-embedded tissue sections (1mm to 4mm) corresponding to the cancer parts of the HE-stained sections, preparing paraffin-embedded tissue cores, preparing breast cancer needle biopsy tissue arrays by utilizing a tissue chip instrument, cutting paraffin into slices, and preparing paraffin-embedded tissue chip of breast cancer needle biopsy tissue. The prepared needle biopsy tissue chip can be applied to molecular medical research of related diseases such as immunocytochemistry, in-situ hybridization, and the like.

The invention discloses a tissue chip receptor blank wax block preparation device, relates to a mould and particularly relates to a mould for the tissue chip, a de-molding device and a using method of the tissue chip receptor blank wax block preparation device. The tissue chip receptor blank wax block preparation device is relatively simple, flexible, relatively convenient to operate, convenient to de-mould and high in de-molding precision. A lifting device fixed on a base is used for jacking a pushing rod on a pushing plate; the pushing rod penetrates through a base pushing hole and a plate pushing hole for jacking an array pore plate; a guide rail base is used for fixing an array column plate through a guide rail, so that the array column plate obtains tensile force produced by gravity force of the guide rail base; the thrust force of the pushing rod and the gravity force of the guide rail base co-act for separating the mould for completing de-molding, so that inconvenience caused by manual de-molding is avoided; meanwhile, the direction of the thrust force and the direction of the tensile force are parallel, so that the de-molding is accurate, the property loss caused by manual de-molding errors is avoided, and the wax block is adjustable in size through a vertical plate.

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.

A movable type printing type method for preparing and repariing the tissue chip includes sticking the chip substrate carrying organic adhesive onto the surface of wax block, slicing, sticking the sliced section onto glass plate, baking, cooling, and putting it in organic solvent to dissolve said adhesive and wax.

The invention discloses a cervical cancer prognosis test method which comprises the following steps sequentially of tissue chip processing, result judgment, determination and the like. According to the method, the SETD4 expression level is tested through immunohistochemistry and with a quantitative PCR method, a relatively normal tissue is found, the uterine cervicitis tissue SETD4 lightness is improved, the tumor tissue is improved remarkably, and the SETD4 expression level can be used for predicating cervical cancer prognosis and taken as a prognostic indicator for predicating the cervical cancer therapeutic effect.

The invention provides a method for making a tissue chip based on one-step molding. The method is characterized in that the tissue chip is made through one-step molding without preparing a donor which is paraffin or an acceptor which is a paraffin tissue block. The method concretely comprises the following steps: 1, taking a mold frame, putting a tissue in the embedding tube of the mold frame in a required size and a required tangent plane, dumping paraffin for embedding, and allowing the paraffin to stand for a period of time to carry out polymerization molding of the paraffin; and 2, taking a paraffin-embedded frame, placing the mold frame having the embedding structure obtained in step 1 in the embedding frame on flat glass, dumping molten paraffin into the paraffin-embedded frame, allowing the paraffin in the porous tubes of the mold frame and the paraffin in the embedded frame to melt, and removing the embedded frame after the melting to complete the tissue embedding. According to the invention, the tissue positions and the embedding accuracy are strictly controlled, and no burrs or discontinuous cracks appear during punching, so the method has the advantages of cheapness, convenience, high efficiency, time saving and labor saving, and can be popularized to each pathology department and relevant research units.

The invention provides a method for making a once-forming tissue chip and a mold applied to the same, belonging to the technical field of tissue chip making and solving the problems of high technical requirements and the like in the traditional tissue chip making process. The method comprises the following steps of: firstly, assembling a mold; secondly, taking a core and filling; thirdly, preheating and injecting wax; fourthly, disassembling; and fifthly, slicing and dyeing. The method can be used for making the once-forming tissue chip, simplify the making flow and has the advantages of simple operation steps, rapidness, economy, easiness of use and the like.

The invention discloses a device for preparing a blank receptive block of a tissue chip with holes. The device comprises a pore plate, columnar fine needles and a detachable rectangular bulkhead which forms a box structure together with the pore plate, wherein, the pore plate is provided with through holes which are regularly arranged in rows; the through holes are T-shaped through holes composedof fine holes and big holes, which are vertically communicated; accordingly, the columnar fine needles are reducing needles which are embedded in and pass through the T-shaped through holes; at leastthe thin part length of one reducing needle is longer than vertical lengths of fine holes and the thick part lengths of the reducing needles are longer than vertical lengths of big holes. The device can effectively determine the axial position of a needle and strictly control the punching depth on the basis of realizing one-step molding; the punching accuracy is easy to control; the holes have noburrs or chap; finished wax block products are not damaged during the production, and the uniformity of punching on the wax block is ensured, so that the production quality of the wax block is ensured and the work efficiency is improved while time and labor are saved.

The invention discloses a method for detecting intratumoral heterogeneity of triple-negative breast cancer. The method solidifies tissue samples from different parts of the same patient on the same tissue chip, and then performs immunohistochemistry on the tissue chip. , immunofluorescence, fluorescence in situ hybridization and other detection. Effectively reduce the cost of experiments, save time and manpower, and also reduce errors caused by batch experiments, improve the consistency of experimental results, and achieve better quality control. This method uses multi-probe combined with fluorescence in situ hybridization technology to evaluate the intratumoral heterogeneity of triple-negative breast cancer, which reduces the dependence on expensive equipment and investment costs, and is easy to be widely promoted. The invention quantitatively detects intratumoral heterogeneity of triple-negative breast cancer, and provides a clear evaluation index for accurately predicting postoperative recurrence and metastasis risk of patients.

A process for preparing paraffin wax embedded tissue microarray chip with oridinary microscope includes such steps as installing special perforating needle and sampling needle to the lens holder of the ordinary optical microscope, putting special paraffin wax block fixing box on object carrier, turning the lens holder and regulating the distance between lens and object carrier while perforating, sample and checking sample, and observing while locating. Its advantages are simple operation and low cost.

The invention discloses a manufacturing method for a tissue chip, which comprises the following steps: the manufacture and perforation of a receptor wax block, the manufacture of the mark of a point to be taken and a donor wax block, the sampling of the donor wax block, recording of array point information, and the processing and slicing of an array wax block; wherein, the manufacture of the mark of the point to be taken and the donor wax block comprises the following steps: (1) when obtaining the materials, a needle is used for perforating at 1 / 3 position of the left upper quadrant of the wax block so as to mark as the origin a of an XY-axis; (2) the a is taken as the origin, and coordinate distances of the point which needs to obtain materials on the horizontal axis (X-axis) and the vertical axis (Y-axis) are respectively measured in a microscope; (3) marking is carried out when returning the corresponding coordinate point on the wax block; (4) the sampling needle is used for perforating and sampling on the parts to be taken out on the donor wax block; (5) the sample is planted accurately in the array wax block (receptor wax block); the manufacturing method of the invention can achieve the material-obtaining of precise point by point to point, thus avoiding ineffective chip deviation caused by heterogeneity of tumors reported before; the manufacturing method can also record point information of each chip precisely which is convenient for test in future, and can establish a tumor tissue chip library with normal series effectively.

The invention discloses application of a GPR115 gene in preparation of an anti-lung cancer drug and a diagnostic kit thereof, and belongs to the technical field of cancer precision medical drugs. Immunohistochemistry proves that the expression of GPR115 in lung cancer is increased through a lung cancer clinical sample tissue chip, and GPR115 is related to prognosis; the influence of down-regulation of GPR115 gene expression by siRNA on biological behaviors such as proliferation and invasion of lung cancer cells is studied through an in-vitro cell experiment, it is found that the specific siRNAsequence can effectively inhibit the expression of GPR115 protein in human lung cancer cell strains H1650 and SPCA, and after the expression of GPR115 is inhibited by siRNA, the proliferation of H1650 and SPCA cells is slowed down, and the cell invasion ability is reduced. The GPR115, as a target for lung cancer gene therapy, can be widely applied to preparation of a diagnostic kit for precisionmedical treatment and treatment of lung cancer with high expression of GPR115.

The invention provides a method for mass production of a tissue or cell chip. The method comprises shaping tissue particles, cultured cells or collected exfoliated cells (such as pleuroperitoneal fluid exfoliated cells) through a shaping device. The tissue particles are obtained through smashing fresh tissue, tissue fixed by formalin or other solvents (such as alcohol), or paraffin-embedded tissueblocks through a tissue blender. The shaped tissue or cells are implanted into a receptor wax mold or are bonded to a metal plate and then are sliced so that a tissue or cell chip is obtained. The method has the advantages of low cost, convenience, rapidity, mass production and variety of shapes, can be used for exfoliated cells of fresh tissue, fixed tissue, paraffin-embedded tissue and cultureor pleuroperitoneal fluid, and can be spread to every pathology and related research unit.