CN103869057B - Needle biopsy tissue chip - Google Patents
Needle biopsy tissue chip Download PDFInfo
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- CN103869057B CN103869057B CN201210538814.1A CN201210538814A CN103869057B CN 103869057 B CN103869057 B CN 103869057B CN 201210538814 A CN201210538814 A CN 201210538814A CN 103869057 B CN103869057 B CN 103869057B
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Abstract
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.
Description
Technical field
The invention belongs to biochip field, it is related to a kind of biochip for preparing microarray punch biopsies chip and preparation method.
Background technology
High density tissue that biochip refers to be coated on the solid supports such as silicon chip, nylon membrane, cell, protein, the high flux microarray of nucleic acid, saccharide and other biological component.Using high flux biochip, hybridized by specific molecular and staining technique, examined samples detected, can accurately, quick obtaining substantial amounts of Molecular Detection information.It is widely used to life science, clinical medicine molecular diagnosis and the field such as biological antibody targeted drug examination and checking at present.
Punch biopsies pathological examination, it has also become one of clinical pathology diagnosis routine examination method.Punch biopsies diameter typically between about 1.0mm ~ 2.0mm, strip of tissue is conventional fixing, dehydration embedding, after paraffin section pathological diagnosis, remaining tissue mass is less.If follow-up, because research needs continuation, routine paraffin wax is cut into slices, and remaining tissue has been readily consumed by.In order to improve the service efficiency of remaining tissue samples after clinical punch biopsies pathological examination, a kind of technology of preparing of high-throughout punch biopsies chip and method are invented.
Still do not prepare the technology of organization chip and the patent of application and document report using punch biopsies at present.
Content of the invention
The present invention is intended to provide a kind of technology and method preparing high-throughout punch biopsies chip.Prepared punch biopsies chip can be used for the SABC of relevant disease, in situ hybridization equimolecular medical research.
The present invention is achieved through the following technical solutions goal of the invention.Collect the aspiration biopsy residue paraffin organization specimen of 42 patients being diagnosed as breast carcinoma through breast puncture biopsy pathological examination, with corresponding routine pathology he stained for guiding, cut he stained cancer partly corresponding paraffin organization bar section (1mm-4mm), prepare paraffin organization core, prepare breast carcinoma punch biopsies array using organization chip instrument, paraffin section prepares the paraffin organization chip of breast carcinoma punch biopsies.
Brief description
Fig. 1 is the schematic diagram of hollow array paraffin mass wax-pattern: wherein, 1: the paraffin hole reserved for 2 breast carcinoma paraffin cores；2: the paraffin hole reserved for 40 breast carcinoma paraffin cores.All apertures are 1.5mm, and pitch of holes is 1mm.
Fig. 2 is the schematic diagram of breast cancer tissue array paraffin mass: wherein, 1: for 2 breast carcinoma paraffin stem stems；2: for 40 breast carcinoma paraffin stem stems.Aspiration biopsy paraffin organization column diameter is 1.5mm, and paraffin organization intercolumniation is 1mm.
Fig. 3 is the schematic diagram of aspiration biopsy paraffin organization chip: due to pathological diagnosis paraffin section, aspiration biopsy paraffin organization post longitudinally by partially sliced excision, organizes post lateral cross section to become the disc shape of part.Wherein, 1: for 2 breast carcinoma paraffin chip interlacing points, simultaneously also can be used as the location determination site of this aspiration biopsy paraffin organization chip；2: for 40 breast carcinoma paraffin chip interlacing points.A diameter of 1.5mm of aspiration biopsy paraffin organization chip interlacing point, breast carcinoma paraffin chip tissue dot spacing is 1mm.3: for the frosted marked region on sperm chip solid phase carrier (adhesion microscope slide).
Specific embodiment
Embodiment: collect the aspiration biopsy residue paraffin organization specimen of 42 patients being diagnosed as breast carcinoma through breast puncture biopsy pathological examination, the paraffin organization chip of preparation breast carcinoma punch biopsies.
1.1 pathological diagnosis: breast puncture biopsy bar, after routine pathology is fixed, is dehydrated embedding, paraffin section dyes, carries out pathological diagnosis.
1.2 specimen are selected: select 42 breast puncture biopsies diagnosis to be diagnosed as the paraffin tissue sections of breast carcinoma, the region (> 1mm of breast carcinoma focus in labelling he stained) as organization chip preparation destination organization bar.
1.3 paraffin cores preparation: with he stained the region of breast carcinoma focus be labeled as guide, with paraffin section knife by corresponding Paraffin tissue block puncturing tissue bar cut off, cutting tissue bar length be 1mm-4mm.Paraffin tissue block is put on the metal heating disc of 70 DEG C of preheatings (paraffin mass section faces down), heating paraffin wax piece of tissue sliced surfaces about 1 minute, melts to Paraffin tissue block superficial paraffin fractions, carefully take off breast carcinoma focus target paraffin organization bar with tweezers.Target paraffin organization bar is put on metal heating disc and continues to be heated to strip of tissue paraffin to melt completely.Carefully the paraffin organization bar of thawing is longitudinally put into (70 DEG C of preheatings) in the plastic syringe of homemade a diameter of 1.5mm.The plastic syringe integral level that will be equipped with target paraffin organization bar is put in the paraffin of a small amount of thawing, to be saturated with paraffin in plastic syringe.Plastic syringe is taken out to be placed in 4 DEG C of cold bench and quickly cools down.Naked eyes observe, to light, the position that can clearly see that plastic syringe inner tissue bar is located, and the position with marking pen tagged tissue bar two ends.Cut off plastic syringe, carefully longitudinally slit plastic syringe (attention tries not to switch to strip of tissue) with paraffin section knife in strip of tissue two ends mark, take out paraffin organization bar, as: paraffin core.
1.4 receptor wax-patterns: the blank paraffin mass of preparation, drill through the hollow receptor wax-pattern that aperture is 1.5mm, such as accompanying drawing 1 using organization chip preparing instrument positioning.
1.5 tissue arrays: according to the design of the paraffin organization chip of breast carcinoma punch biopsies, altogether corresponding blank well site will be placed in hollow receptor wax-pattern one by one by 42 paraffin cores, on the basis of being flushed by array surface, such as accompanying drawing 2.
1.6 arrays merge: tissue array is placed in drying baker, adjusts drying baker temperature and time it is ensured that all tissue paraffin cores and receptor wax-pattern fully merge and paraffin core does not shift deformation.
1.7 paraffin sections: using paraffin slicing machine microsection manufacture paraffin section, piece thickness 4um.Paraffin section is tiled and adheres on microscope slide, such as accompanying drawing 3.Perform labelling in microscope slide marked region.
The paraffin organization chip chip manufacturing of 1.8 breast carcinoma punch biopsies finishes.In conjunction with the experimental molecule investigative technique such as SABC, in situ hybridization, the paraffin organization chip of prepared breast carcinoma punch biopsies can be used for the molecular medicine research of breast carcinoma.
Claims (4)
1. a kind of preparation method of punch biopsies chip it is characterised in that: comprise the following steps:
The first step, pathological diagnosis: breast puncture biopsy bar, after routine pathology is fixed, is dehydrated embedding, paraffin section dyes, carries out pathological diagnosis；
Second step, specimen is selected: selects 42 breast puncture biopsies diagnosis to be diagnosed as the paraffin tissue sections of breast carcinoma, the destination organization bar that in labelling he stained, the region of breast carcinoma focus is prepared as organization chip；
3rd step, paraffin core prepare: with he stained the region of breast carcinoma focus be labeled as guide, with paraffin section knife by corresponding Paraffin tissue block puncturing tissue bar cut off, cutting tissue bar length be 1mm-4mm；Paraffin tissue block section is faced down be put on 70 DEG C of metal heating discs preheating and heat, heating paraffin wax piece of tissue sliced surfaces about 1 minute, paraffin melting to Paraffin tissue block superficial, take off focus target paraffin organization bar with tweezers；Target paraffin organization bar is put on metal heating disc and continues to be heated to strip of tissue paraffin to melt completely；In the self-control plastic syringe of a diameter of 1.5mm paraffin organization bar of thawing longitudinally being put into 70 DEG C of preheatings；The plastic syringe integral level that will be equipped with target paraffin organization bar is put in the paraffin of a small amount of thawing, to be saturated with paraffin in plastic syringe；Plastic syringe is taken out to be placed in 4 DEG C of cold bench and quickly cools down；Naked eyes observe, to light, the position that can clearly see that plastic syringe inner tissue bar is located, with the position at marking pen tagged tissue bar two ends；Cut off plastic syringe with paraffin section knife in strip of tissue two ends mark, longitudinally cut plastic syringe open, take out paraffin organization bar, as: paraffin core；
4th step, receptor wax-pattern: the blank paraffin mass of preparation, drill through, using organization chip preparing instrument positioning, the hollow receptor wax-pattern that aperture is 1.5mm；
5th step, tissue array: according to the design of the paraffin organization chip of breast carcinoma punch biopsies, paraffin core is placed in hollow receptor wax-pattern corresponding blank well site one by one, on the basis of flushing by array surface；
6th step, array merges: tissue array is placed in drying baker, adjusts drying baker temperature and time it is ensured that all tissue paraffin cores and receptor wax-pattern fully merge and paraffin core does not shift deformation；
7th step, paraffin section: using paraffin slicing machine microsection manufacture paraffin section, piece thickness 4um；By in paraffin section tiling and adhesion microscope slide；Perform labelling in microscope slide marked region；
8th step, the paraffin organization chip of breast carcinoma punch biopsies makes and finishes.
2. the preparation method of a kind of punch biopsies chip according to claim 1, it is characterized in that: described punch biopsies chip is including but not limited to breast carcinoma punch biopsies specimen, can also be the punch biopsies specimen of other malignant tumor, or the punch biopsies specimen of benign tumor and non-tumor.
3. the preparation method of a kind of punch biopsies chip according to claim 1, it is characterized in that: in clinical aspiration biopsy paraffin organization, the acquisition modes of destination organization with operation sequence are: first by destination organization, on corresponding he dyed paraffin is cut into slices, pathological diagnosis positioning, positioning cut off destination organization bar, then heating and melting obtains destination organization bar, and destination organization bar is put into cooling plasticity in the suitable plastic syringe of internal diameter, prepare paraffin stem stem.
4. a kind of punch biopsies chip according to claim 1 preparation method it is characterised in that: punch biopsies chip tissue spot diameter be 1.0mm-1.5mm between；The spacing of organization chip point is 1mm.
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| CN201210538814.1A CN103869057B (en) | 2012-12-13 | 2012-12-13 | Needle biopsy tissue chip |
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| CN201210538814.1A CN103869057B (en) | 2012-12-13 | 2012-12-13 | Needle biopsy tissue chip |
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| CN103869057A CN103869057A (en) | 2014-06-18 |
| CN103869057B true CN103869057B (en) | 2017-01-18 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108181148A (en) * | 2017-12-25 | 2018-06-19 | 天津市海河医院 | The tissue tabletting of CT Guided Percutaneous Transthoracic Needle Aspiration Biopsies and field evaluation method |
| CN108387419A (en) * | 2018-02-09 | 2018-08-10 | 青岛锴锦医疗器械有限公司 | A kind of novel environment friendly quick bio tissue treatment liquid |
| CN108593380B (en) * | 2018-04-25 | 2021-05-18 | 复旦大学附属中山医院 | Mass production manufacturing method of tissue chips |
| CN110208056B (en) * | 2019-06-06 | 2021-08-31 | 厦门大学附属第一医院 | Method for manufacturing tissue chip of gastric cancer HER-2 FISH |
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| CN1356553A (en) * | 2001-09-13 | 2002-07-03 | 陕西超英生物医学研究开发有限公司 | Tissue microarray biochip |
| US6696271B2 (en) * | 2001-08-23 | 2004-02-24 | The Regents Of The University Of California | Frozen tissue microarray technology for analysis of RNA, DNA, and proteins |
| CN101231282A (en) * | 2007-01-23 | 2008-07-30 | 北京市农林科学院 | Organization chip for researching functional genome as well as preparation method and application thereof |
| CN101319971A (en) * | 2008-07-01 | 2008-12-10 | 南通大学 | Preparation method of paraffin organization chip |
| CN101738338A (en) * | 2009-12-29 | 2010-06-16 | 李从善 | Method for manufacturing tissue array paraffin block |
| CN102798562A (en) * | 2012-08-02 | 2012-11-28 | 王虎 | Paraffin texture chip preparation method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| PT1756545E (en) * | 2004-06-18 | 2011-10-24 | Covance Inc | Frozen tissue microarray |
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| US6696271B2 (en) * | 2001-08-23 | 2004-02-24 | The Regents Of The University Of California | Frozen tissue microarray technology for analysis of RNA, DNA, and proteins |
| CN1356553A (en) * | 2001-09-13 | 2002-07-03 | 陕西超英生物医学研究开发有限公司 | Tissue microarray biochip |
| CN101231282A (en) * | 2007-01-23 | 2008-07-30 | 北京市农林科学院 | Organization chip for researching functional genome as well as preparation method and application thereof |
| CN101319971A (en) * | 2008-07-01 | 2008-12-10 | 南通大学 | Preparation method of paraffin organization chip |
| CN101738338A (en) * | 2009-12-29 | 2010-06-16 | 李从善 | Method for manufacturing tissue array paraffin block |
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