CN1253716C - Tissue chip used for tumour early stage diagnosis and preparation device - Google Patents
Tissue chip used for tumour early stage diagnosis and preparation device Download PDFInfo
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- CN1253716C CN1253716C CN 200410022818 CN200410022818A CN1253716C CN 1253716 C CN1253716 C CN 1253716C CN 200410022818 CN200410022818 CN 200410022818 CN 200410022818 A CN200410022818 A CN 200410022818A CN 1253716 C CN1253716 C CN 1253716C
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Abstract
The present invention relates to a tissue chip. The present invention selects paraffin tissue specimens of common malign tumors with complete clinical data and pathological characteristics, such as lung cancer, nasopharynx cancer, esophagus cancer, stomach cancer, large intestine cancer, liver cancer, galactophore cancer, cervix cancer, etc. The tissue specimens in precancerous lesion and corresponding normal tissue specimens are sliced up, dyed, marked and positioned; a piece of lattice array mould paper is designed and pasted on the surface of a receptor paraffin block for guiding the preparation of receptor holes; paraffin blocks with tissue core strips are prepared by using simple and practical perforating needles and tissue puncture needles. The present invention is a tissue chip product which comprises cancer tissues, tissues in precancerous lesion and corresponding normal tissues. Proved by hybridization in situ and immunohistochemical assay, the expressions of relevant gene mRNA and proteins in the tissue chip are consistent with the result of a conventional test; cell form is clear, even and identical, and the tissue chip does not have the shedding of tissue points. Each specimen in the tissue chip has corresponding clinical and pathology data; the tissue chip can be used for testing the expression differences of various target genes in a large number of specimens and polytype cancer tissues, and provides a high-flux tool for selecting molecule markers for the early diagnosis and the prediction prognosis of tumors.
Description
Technical field:
The present invention relates to the biological products that tissue detection is used, be specifically related to a kind of organization chip.
Background technology:
Organization chip claims micro-array tissue (Tissue chip again, Tissue Microarray TMA,) be that many donor tissue rearrange on acceptor wax block with micro-array, to acceptor wax block section and be transferred on the microslide, thisly comprise that the sections of hundreds of tissues can be used for various expressed in situ researchs.Organization chip often utilizes the micro-array tissue instrument to carry out the puncture of donor wax stone tissue and the making of acceptor wax block receptor hole.Tumor tissues chip, normal structure chip, single or compound organization chip etc. are arranged in the market, but can not be used for high flux screening diagnosing early malignant tumor and predict prognosis molecular marker.Therefore, press for to make and a kind ofly can be used for detecting various genes of interest in the large sample amount, the organization chip of the differential expression in the polytype cancerous tissue is to solve the problem that the current organization chip exists.
Summary of the invention:
The present invention is intended to develop a kind of novel organization chip, to solve the problem that can not be used for high flux screening diagnosing early malignant tumor and predict prognosis molecular marker that the current organization chip exists.
The present invention is achieved by the following technical solutions.
The making that is used for the organization chip of early diagnosis of tumor and predict prognosis molecular marker screening comprises the making of the making of sample preparations, acceptor paraffin mass, organization chip paraffin mass and organization chip section.The preparation of sample will be selected the paraffin organization sample of paraffin organization sample, precancerous lesion and the related normal tissue of the common cancer with complete clinical and pathological characters data; The making of acceptor paraffin mass will be by making organization chip density and organize the size of core bar diameter, is affixed on the acceptor wax block surface with the mould paper of the lattice array of corresponding specification; The making of organization chip paraffin mass is puncture needle to be worn get tissue from each sample, releases circular paraffin organization core bar and moves into corresponding being subjected to the paraffin body hole from the inner core of this puncture needle.Making the used puncture needle of organization chip section is a kind of lumbar puncture needle that circular sharp front end is arranged, the inner core of puncture needle is longer than about puncture needle 1.0cm, the external diameter of the perforating needle of acceptor paraffin mass punching usefulness is consistent with puncture needle inner core size, and perforating needle and tissue penetration shank bag are convenient to hold the overcoat of pin operation and restriction paracentesis depth.
Be described in further detail the present invention below in conjunction with accompanying drawing.
Description of drawings:
Fig. 1 is the perforating needle synoptic diagram; Wherein: 1, punching needle tubing 2, perforating needle overcoat
Fig. 2 is puncture needle and nook closing member synoptic diagram; Wherein: 3, puncture needle tubing 4, puncture needle overcoat 5, nook closing member
Fig. 3 is the pattern paper of 448 dot matrix;
Fig. 4 organizes the core bar for puncture;
Fig. 5 is 448 acceptor wax blocks of organizing dot matrix;
Fig. 6 is 448 HE sections of organizing dot matrix;
Fig. 7 is HE dyeing 4 * nasopharyngeal carcinoma organization chip cellular morphology figure;
Fig. 8 is a SABC, P53+4 * nasopharyngeal carcinoma organization chip cellular morphology figure;
Fig. 9 is in situ hybridization, BRD7+4 * nasopharyngeal carcinoma organization chip cellular morphology figure.
Organization chip of the present invention is the novel organization chip product that comprises cancerous tissue, precancerous lesion tissue and related normal tissue.Each sample all has corresponding clinical and pathological data in this chip, the concrete method for making of this chip is: (1) sample preparations and purpose are organized HE stained location: select the lung cancer with complete clinical and pathological characters data respectively from the tissue specimen storehouse, nasopharyngeal carcinoma, the cancer of the esophagus, cancer of the stomach, colorectal cancer, liver cancer, breast cancer, each 300 example of the paraffin organization sample of common cancers such as cervical carcinoma, precancerous lesion 200 example and related normal tissue 100 examples, conventional dehydration, paraffin embedding, section, HE dyeing, microscopically confirms to organize correctness, at microscopically purpose tissue site in cutting into slices is carried out the mark location then.(2) making of acceptor paraffin mass: according to making organization chip density and the size of organizing core bar diameter, utilizing the mould paper (Fig. 3) of corresponding specification lattice array to be affixed on the acceptor wax block surface guides receptor hole to make, cause paraffin collapse around the acceptor wax block hole when preventing to punch, acceptor wax block is positioned over 37 ℃ of preheating 30min before punching.(3) making of organization chip paraffin mass: utilize the representative tissue regions of HE section sign to locate site of puncture on the donor paraffin mass; Puncture needle is worn from each sample and is got tissue, uses with the supporting inner core of this puncture needle and releases circular paraffin organization core bar (Fig. 4) and move into corresponding being subjected in the paraffin body hole.After treating that whole receptor hole are filled up tissue, acceptor wax block places 37 ℃, and 30min gently presses wax stone donor tissue face with microslide, makes whole donor tissue on same plane.(4) making of organization chip section: with on the cycle type microtome first to the organization chip paraffin mass rough lumber repair sheet, treat that all receptor tissue's dot matrix are all behind a plane, utilizing the paraffin adhesive tape to shift backup system cuts into slices to the acceptor paraffin mass, thickness is that the adhesive tape histotomy of 5 μ m is pasted on the microslide of adhering substrate, crosslinked 1min under the uviol lamp, dissolving of adhesive tape lysate and stripping tape, aquation can conventionally dewax, as the need long preservation, section is immersed fast and is dissolved in the paraffin, slice surface is carried out again covered thin layer paraffin, the section that disposes can be standby-20 ℃ of long term storage.(5) perforating needle and the tissue penetration pin of the section of making organization chip are to utilize the lumbar puncture needle transformation of different size to form, and clip lumbar puncture needle tip side tangent plane, are processed into the tissue penetration pin (Fig. 2) of circular sharp front end; The external diameter of perforating needle (1) is consistent with puncture needle inner core (5) size, and perforating needle and tissue penetration shank bag are convenient to hold pin operation and restriction paracentesis depth by overcoat (2,4), and the inner core of tissue penetration pin (5) is longer than about puncture needle (3) 1.0cm.
The quality evaluation of organization chip practical application: in situ hybridization and SABC detect related gene mRNA and the expression of albumen in organization chip, the result who detects tissue specimen with the conventional sense method is consistent, and inorganization chip point obscission, cellular morphology is clear, uniformity.This organization chip is the same with conventional organization chip, energy and histochemistry, and immunohistochemistry, in situ hybridization, original position PCR, technology such as original position RT-PCR and cDNA chip combine, and are used for gene functional research.In addition, this novel high flux malignant tumor tissue chip with complete clinical and pathological data can be extensive, detect relevant genes of interest DNA cloning in the tissue in position apace, mRNA expresses the relation of abundance and protein expression level and clinical pathologic characteristic, and the molecular marker for screening early diagnosis of tumor and prognosis prediction provides a kind of novel high flux instrument veritably.
Embodiment:
Embodiment: nasopharyngeal carcinoma organization chip and making thereof
(1) sample preparations and purpose are organized HE stained location: select nasopharyngeal carcinoma paraffin organization sample 250 examples with complete clinical and pathological characters data respectively from the tissue specimen storehouse, precancerous lesion 150 examples and corresponding normal nasopharyngeal are organized 48 examples, conventional dehydration, paraffin embedding, section, HE dyeing, microscopically confirms to organize correctness, and microscopically carries out the mark location to purpose tissue in the section.(2) making of acceptor paraffin mass: making and organizing diameter is the 1.0mm size, comprise nasopharyngeal carcinoma, precancerous lesion and normal nasopharyngeal are organized the organization chip of totally 448 samples, utilizing the mould paper (Fig. 3) of 448 interlacing point arrays to be affixed on the acceptor wax block surface guides receptor hole to make, cause paraffin collapse around the acceptor wax block hole when preventing to punch, acceptor wax block is positioned over 37 ℃ of preheating 30min before punching.(3) making of organization chip paraffin mass: utilize the representative tissue regions of HE section sign to locate site of puncture on the donor paraffin mass; Puncture needle is worn from each sample and is got tissue, uses to release circular paraffin organization core bar (Fig. 4) with the supporting inner core of puncture needle and move in the corresponding acceptor wax block hole.After treating that whole receptor hole are filled up tissue, acceptor wax block places 37 ℃, and 30min gently presses wax stone donor tissue face to make whole donor tissue on same plane with microslide.(4) making of organization chip section: with on the cycle type microtome first to the organization chip paraffin mass rough lumber repair sheet, treat that all receptor tissue's dot matrix are all behind a plane, utilizing the paraffin adhesive tape to shift backup system cuts into slices to the acceptor paraffin mass, thickness is that the adhesive tape histotomy of 5 μ m is pasted on the microslide of adhering substrate, crosslinked 1min under the uviol lamp, dissolving of adhesive tape lysate and stripping tape, aquation can conventionally dewax, as the need long preservation, section is immersed fast and is dissolved in the paraffin, slice surface is carried out again covered thin layer paraffin, the section that disposes can be standby-20 ℃ of long term storage.(5) preparation of perforating needle and tissue penetration pin: perforating needle (external diameter 1mm) and tissue penetration pin (internal diameter 1mm) are to utilize the transformation of different size lumbar puncture needle to form, the perforating needle external diameter is consistent with the puncture needle inner core, clip the distolateral tangent plane of needle tip, be processed into circular sharp front end; Perforating needle and tissue penetration shank bag are convenient to hold pin operation and restriction paracentesis depth by overcoat, and the inner core of tissue penetration pin is longer than about puncture needle 1.0cm.(6) quality evaluation of nasopharyngeal carcinoma organization chip practical application: in situ hybridization and SABC detect BRD7 gene mRNA and the expression of P53 albumen in organization chip, the result who detects tissue specimen with the conventional sense method is consistent, and inorganization chip point obscission, cellular morphology clear (Fig. 7-9).
Claims (3)
1, a kind of organization chip that is used for early diagnosis of tumor, the making of this chip comprise the making of making, organization chip paraffin mass and the organization chip section of sample preparations, acceptor paraffin mass, it is characterized in that:
(1) preparation of sample: the paraffin organization sample that select paraffin organization sample, precancerous lesion and the related normal tissue of common cancer with complete clinical and pathological characters data, tissue specimen is through routine dehydration, paraffin embedding, section, HE dyeing, microscopically confirms to organize correctness, at microscopically purpose tissue site in cutting into slices is carried out the mark location then;
(2) making of acceptor paraffin mass: by density of making organization chip and the size of organizing core bar diameter, mould paper with the lattice array of corresponding specification is affixed on the acceptor wax block surface, cause paraffin collapse around the acceptor wax block hole when preventing to punch, acceptor wax block is positioned over 37 ℃ of preheating 30min before punching, punch then;
(3) making of organization chip paraffin mass: puncture needle is worn from each sample and is got tissue, release circular paraffin organization core bar and move into corresponding being subjected to the paraffin body hole from the inner core of this puncture needle, after treating that whole receptor hole are filled up tissue, acceptor wax block places 37 ℃, 30min, gently press wax stone donor tissue face with microslide, make whole donor tissue on same plane;
(4) making of organization chip section: with the cycle type microtome first to the organization chip paraffin mass rough lumber repair sheet, treat that all receptor tissue's dot matrix are all behind a plane, utilizing the paraffin adhesive tape to shift backup system cuts into slices to the acceptor paraffin mass, thickness is that the adhesive tape histotomy of 5 μ m is pasted on the microslide of adhering substrate, crosslinked 1min under the uviol lamp, dissolving of adhesive tape lysate and stripping tape can conventional dewaxing aquations.
2, the organization chip that is used for early diagnosis of tumor as claimed in claim 1 is characterized in that: as the need long preservation, section is immersed fast and is dissolved in the paraffin, makes slice surface cover thin layer paraffin again, and the section that disposes can be standby-20 ℃ of long term storage.
3, a kind of utensil of making the described organization chip of claim 1, it is characterized in that: making the used puncture needle of organization chip paraffin mass is a kind of lumbar puncture needle that circular sharp front end is arranged, the inner core of puncture needle (5) is longer than puncture needle (3) 1.0cm, the external diameter of the perforating needle (1) of acceptor paraffin mass punching usefulness is consistent with puncture needle inner core (5) size, and perforating needle and tissue penetration shank bag are convenient to hold the overcoat (2,4) of pin operation and restriction paracentesis depth.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410022818 CN1253716C (en) | 2004-01-08 | 2004-01-08 | Tissue chip used for tumour early stage diagnosis and preparation device |
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| CN 200410022818 CN1253716C (en) | 2004-01-08 | 2004-01-08 | Tissue chip used for tumour early stage diagnosis and preparation device |
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| CN1556413A CN1556413A (en) | 2004-12-22 |
| CN1253716C true CN1253716C (en) | 2006-04-26 |
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Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4477575B2 (en) * | 2005-12-14 | 2010-06-09 | 株式会社日立製作所 | Gene set used for colorectal cancer testing |
| CN1920558B (en) * | 2005-12-27 | 2013-06-05 | 胡苹 | Wax module for organizing chip array |
| CN1967196B (en) * | 2006-09-14 | 2010-05-12 | 绵竹市人民医院 | Simplified thin-layer liquid-based cytology cell smeard preparation method |
| CN101042316B (en) * | 2007-04-30 | 2011-01-26 | 孙爱静 | Tumor displace mimetism organization chip |
| CN101921858B (en) * | 2010-08-23 | 2013-08-28 | 广州益善生物技术有限公司 | Liquid phase chip for detecting breast cancer prognosis-related gene mRNA expression level |
| CN102466729B (en) * | 2010-11-05 | 2015-06-17 | 北京工业大学 | Method for screening tumor specificity target and targeting ligand based on tissue chip |
| CN102676650B (en) * | 2011-03-09 | 2015-08-05 | 中国医学科学院肿瘤研究所 | The application of detection by quantitative in esophageal squamous cell carcinoma Index for diagnosis of CPT1A gene or albumen |
| CN103376321B (en) * | 2012-04-13 | 2015-02-11 | 朱有凯 | Development and application of antibody gene chip |
| US20140024024A1 (en) * | 2012-07-17 | 2014-01-23 | General Electric Company | Methods of detecting dna, rna and protein in biological samples |
| CN103969096B (en) * | 2014-04-23 | 2016-06-01 | 杭州电子科技大学 | Automatically device and the method thereof of cell block is prepared |
| KR101743283B1 (en) * | 2015-08-31 | 2017-06-02 | 재단법인 의약바이오컨버젼스연구단 | Lung window apparatus based on micro-suction for in vivo microscopic imaging of lung tissue and method for obtaining image using the same |
| CN111088216A (en) * | 2019-12-16 | 2020-05-01 | 齐妍 | Method for separating and detecting cells by paraffin-embedded tissue |
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