CN103869057A

CN103869057A - Needle biopsy tissue chip

Info

Publication number: CN103869057A
Application number: CN201210538814.1A
Authority: CN
Inventors: 郜恒骏; 张可浩; 张新; 盛海辉; 颜虹; 张宇
Original assignee: SHANGHAI OUTDO BIOTECH CO Ltd
Current assignee: SHANGHAI OUTDO BIOTECH CO Ltd
Priority date: 2012-12-13
Filing date: 2012-12-13
Publication date: 2014-06-18
Anticipated expiration: 2032-12-13
Also published as: CN103869057B

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

Aspiration biopsy organization chip

Technical field

The invention belongs to biochip field, relate to a kind of biochip and preparation method for the preparation of microarray aspiration biopsy organization chip.

Background technology

Biochip refers to the high flux microarray of the high density tissue, cell, protein, nucleic acid, carbohydrate and the other biological component that are coated on the solid support such as silicon chip, nylon membrane.Utilize high flux biochip, hybridize and staining technique by specific molecular, examined samples are detected, can be accurately, a large amount of Molecular Detection information of quick obtaining.The fields such as life science, clinical medicine molecular diagnosis and the examination of biological antibody targeted drug and checking are widely used at present.

Aspiration biopsy histopathological examination, has become one of clinicopathologia diagnosis routine inspection method.Aspiration biopsy organizes diameter generally between about 1.0mm ~ 2.0mm, and after conventional fixing, the dehydration embedding of strip of tissue, paraffin section pathological diagnosis, remaining tissue mass still less.If follow-up because research need to continue routine paraffin wax section, residue tissue is easy to run out of.In order to improve the service efficiency that remains tissue samples after clinical aspiration biopsy histopathological examination, technology of preparing and the method for having invented a kind of high-throughout aspiration biopsy organization chip.

Still do not utilize at present patent and the bibliographical information of the application of aspiration biopsy tissue preparation organization chip.

Summary of the invention

The present invention aims to provide a kind of technology and method of preparing high-throughout aspiration biopsy organization chip.Prepared aspiration biopsy organization chip can be used for SABC, the medical research of in situ hybridization equimolecular of relevant disease.

The present invention is achieved through the following technical solutions goal of the invention.Collect 42 examples are diagnosed as the patient of breast cancer aspiration biopsy residue paraffin organization sample through breast puncture biopsy pathological examination, take corresponding routine pathology HE stained as guide, cut paraffin organization bar section (1mm-4mm) corresponding to HE stained cancer kitchen range part, prepare paraffin organization core, utilize organization chip instrument to prepare breast cancer aspiration biopsy tissue array, paraffin section is prepared the paraffin organization chip of breast cancer aspiration biopsy tissue.

Accompanying drawing explanation

Fig. 1 is the schematic diagram of hollow array paraffin mass wax-pattern: wherein, and 1: be the 2 reserved paraffin holes of routine breast cancer paraffin core; 2: be the 40 reserved paraffin holes of routine breast cancer paraffin core.All apertures are 1.5mm, and pitch of holes is 1mm.

The schematic diagram of Tu2Shi breast cancer tissue array paraffin mass: wherein, 1: be 2 routine breast cancer paraffin stem stems; 2: be 40 routine breast cancer 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: be 2 routine breast cancer paraffin chip interlacing points, also can be used as the location determination site of this aspiration biopsy paraffin organization chip simultaneously; 2: be 40 routine breast cancer paraffin chip interlacing points.The diameter of aspiration biopsy paraffin organization chip interlacing point is 1.5mm, and it is 1mm that breast cancer paraffin chip is organized dot spacing.3: be the frosted marked region on sperm chip solid phase carrier (adhesion microslide).

Embodiment

Embodiment: collection 42 examples are diagnosed as the patient's of breast cancer aspiration biopsy residue paraffin organization sample, the paraffin organization chip of preparation breast cancer aspiration biopsy tissue through breast puncture biopsy pathological examination.

1.1 pathological diagnosis: breast puncture biopsy bar is fixed, dewatered after embedding, paraffin section dyeing through routine pathology, carries out pathological diagnosis.

1.2 samples are selected: the paraffin organization of selecting 42 routine breast puncture biopsy pathology diagnosis to be diagnosed as breast cancer is cut into slices, the destination organization bar that in mark HE stained, prepare as organization chip in the region (>1mm) of breast cancer focus.

1.3 paraffin cores preparations: with the guide that is labeled as in the region of breast cancer focus in HE stained, the puncture strip of tissue in corresponding Paraffin tissue block is cut off with paraffin section cutter, cutting strip of tissue length is 1mm-4mm.Paraffin tissue block is put on the metal heating disc of 70 ℃ of preheatings (paraffin mass sliced surfaces down), heating Paraffin tissue block sliced surfaces approximately 1 minute, to the shallow table paraffin of Paraffin tissue block partial melting, carefully takes off breast cancer focus target paraffin organization bar with tweezers.Target paraffin organization bar is put in and on metal heating disc, continues to be heated to strip of tissue paraffin and melt completely.Carefully the paraffin organization bar of thawing is longitudinally put into the plastics tubule that homemade diameter is 1.5mm (70 ℃ of preheatings).The plastics tubule integral level that target paraffin organization bar is housed is put into the paraffin of a small amount of thawing, to soak full paraffin in plastics tubule.The taking-up of plastics tubule is placed on 4 ℃ of cold platforms cooling fast.Naked eyes are observed and can clearly be seen the position at plastics tubule inner tissue's bar place light, and with the position at marking pen tagged tissue bar two ends.With paraffin section cutter, at strip of tissue two ends mark cut-out plastics tubule, the careful plastics tubules (attention tries not to switch to strip of tissue) that longitudinally cut, take out paraffin organization bar, are: paraffin core.

1.4 acceptor wax-patterns: prepare blank paraffin mass, utilize organization chip preparing instrument to locate and drill through aperture the hollow acceptor wax-pattern that is 1.5mm, as accompanying drawing 1.

1.5 tissue arrays: according to the design of the paraffin organization chip of breast cancer aspiration biopsy tissue, 42 routine paraffin cores are placed to corresponding blank well site in hollow acceptor wax-pattern one by one altogether, flush as benchmark, as accompanying drawing 2 take array surface.

1.6 arrays merge: tissue array is placed in to drying box, regulates drying box temperature and time, guarantee paraffin core and acceptor wax-pattern fully merges and paraffin core is not shifted distortion in a organized way.

1.7 paraffin sections: utilize paraffin slicing machine microsection manufacture paraffin section, the thick 4um of sheet.By on paraffin section tiling and adhesion microslide, as accompanying drawing 3.Perform mark at microslide marked region.

The paraffin organization chip chip manufacturing of 1.8 breast cancer aspiration biopsy tissues is complete.The experiment such as binding immunoassay group, hybridization histochemistry molecular studies technology, the paraffin organization chip of prepared breast cancer aspiration biopsy tissue can be used for the molecular medicine research of breast cancer.

Claims

1. a preparation method for aspiration biopsy organization chip, is characterized in that: comprise the following steps:

The first step, pathological diagnosis: breast puncture biopsy bar is fixed, dewatered after embedding, paraffin section dyeing through routine pathology, carries out pathological diagnosis.

Second step, sample is selected: the paraffin organization of selecting 42 routine breast puncture biopsy pathology diagnosis to be diagnosed as breast cancer is cut into slices, the destination organization bar that in mark HE stained, prepare as organization chip in the region (>1mm) of breast cancer focus.

The 3rd step, paraffin core preparation: with the guide that is labeled as in the region of breast cancer focus in HE stained, the puncture strip of tissue in corresponding Paraffin tissue block is cut off with paraffin section cutter, cutting strip of tissue length is 1mm-4mm.Paraffin tissue block is put on the metal heating disc of 70 ℃ of preheatings (paraffin mass sliced surfaces down), heating Paraffin tissue block sliced surfaces approximately 1 minute, to the shallow table paraffin of Paraffin tissue block partial melting, carefully takes off breast cancer focus target paraffin organization bar with tweezers.Target paraffin organization bar is put in and on metal heating disc, continues to be heated to strip of tissue paraffin and melt completely.Carefully the paraffin organization bar of thawing is longitudinally put into the plastics tubule that homemade diameter is 1.5mm (70 ℃ of preheatings).The plastics tubule integral level that target paraffin organization bar is housed is put into the paraffin of a small amount of thawing, to soak full paraffin in plastics tubule.The taking-up of plastics tubule is placed on 4 ℃ of cold platforms cooling fast.Naked eyes are observed and can clearly be seen the position at plastics tubule inner tissue's bar place light, and with the position at marking pen tagged tissue bar two ends.With paraffin section cutter, at strip of tissue two ends mark cut-out plastics tubule, the careful plastics tubules (attention tries not to switch to strip of tissue) that longitudinally cut, take out paraffin organization bar, are: paraffin core.

The 4th step, acceptor wax-pattern: prepare blank paraffin mass, utilize organization chip preparing instrument to locate and drill through aperture the hollow acceptor wax-pattern that is 1.5mm, as accompanying drawing 1.

The 5th step, tissue array: according to the design of the paraffin organization chip of breast cancer aspiration biopsy tissue, 42 routine paraffin cores are placed to corresponding blank well site in hollow acceptor wax-pattern one by one altogether, flush as benchmark, as accompanying drawing 2 take array surface.

The 6th step, array merges: tissue array is placed in to drying box, regulates drying box temperature and time, guarantee paraffin core and acceptor wax-pattern fully merges and paraffin core is not shifted distortion in a organized way.

The 7th step, paraffin section: utilize paraffin slicing machine microsection manufacture paraffin section, the thick 4um of sheet.By on paraffin section tiling and adhesion microslide, as accompanying drawing 3.Perform mark at microslide marked region.

The 8th step, the paraffin organization chip chip manufacturing of breast cancer aspiration biopsy tissue is complete.The experiment such as binding immunoassay group, hybridization histochemistry molecular studies technology, the paraffin organization chip of prepared breast cancer aspiration biopsy tissue can be used for the molecular medicine research of breast cancer.

2. the preparation method of a kind of aspiration biopsy organization chip according to claim 1, it is characterized in that: described aspiration biopsy organization chip is including but not limited to breast cancer aspiration biopsy tissue specimen, also can be the aspiration biopsy tissue specimen of other malignant tumours, or the aspiration biopsy tissue specimen of benign tumour and non-tumour.

3. the preparation method of a kind of aspiration biopsy organization chip according to claim 1, it is characterized in that: in clinical aspiration biopsy paraffin organization, the obtain manner of destination organization and running program are: first by destination organization pathological diagnosis location, orientation amputate destination organization bar on corresponding HE dyeing paraffin section, then heating and melting is obtained destination organization bar, and destination organization bar is put into the cooling plasticity of plastics tubule that internal diameter is suitable, prepare paraffin stem stem.

4. the preparation method of a kind of aspiration biopsy organization chip according to claim 1, is characterized in that: aspiration biopsy organization chip interlacing point diameter is 1.5mm but is not limited to 1.5mm, can be between 1.0mm-2.0mm; The spacing of organization chip point is 1mm.