CN1356553A - Tissue microarray biochip - Google Patents
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- CN1356553A CN1356553A CN 01128783 CN01128783A CN1356553A CN 1356553 A CN1356553 A CN 1356553A CN 01128783 CN01128783 CN 01128783 CN 01128783 A CN01128783 A CN 01128783A CN 1356553 A CN1356553 A CN 1356553A
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Abstract
A tissue microarray biochiop is mechanically prepared through integrating tens-thousands doner paraffine samples of tissue or cell in an empty receptor paraffine block, slicing, and transferring the microarray section onto glass chip for further analysis.
Description
One. technical field
The present invention relates to a kind of biochip in biomedical research field, further relate to a kind of tissue microarray biochip that is used for multiple biomedical research purpose that adopts the mechanization preparation.
Two. background technology
Correct diagnosis is most important for treatment of diseases.Along with the progress and the development of medical science, new diagnostic method continues to bring out.But up to now, the most direct, reliable diagnostic method still is traditional pathology method.Its basis for estimation mainly is cellular morphology and the dyeing characteristic under the light microscopic, can define disease in view of the above, classification, somatotype, diagnosis, guidance treatment and evaluate its prognosis etc.But the diagnosis basis that traditional pathology method provides and the information of molecular level are very limited.As some breast cancer, its cell surface has the estrogen receptor of high expressed, so be a kind of typical estrogen-dependent tumour, the treatment of prompting antiestrogenic medicine effectively, but traditional pathology method is difficult to find the variation of this molecular level, also is like this for change of specific antigen, carcinomebryonic antigen and the gene thereof of tumour cell etc.The new development of molecular medicine aspect is readily solved this problem, as immunohistochemical staining, in situ hybridization and original position PCR etc.
The diagnosis of disease is all closely related with histiocytic gene and the variation of relevant molecule thereof with treatment, and this molecule variation just has clinical value when being ubiquitous.In fact, the evaluation for a molecular marker needs hundreds of tissue specimens to thousands of different stages of development.Yet traditional pathology method and be based upon immunohistochemistry on its basis, the tissue specimen that in situ hybridization and original position PCR etc. is detected in most of the cases all is single, that be dispersed in, accidental, isolated.If carry out correct evaluation to a molecular marker then needs expensive human and material resources and financial resources, and be a very very long and complicated process.
In order to solve this difficult problem, many scientists have carried out research a large amount of, once reported as people such as Battifora that (LabInvest 1986,55:244-248) will dewax the dehydration tissue specimen manually be rolled into sausage shaped, combination at random, paraffin is embedding again, and conventional section detects a plurality of tissue specimens on same the slide with immunohistochemical method then.Next year, people such as Wan have reported a kind of improved method (Wan et al, J Immumol Meth1987 103:121-129), this method is with paraffin-embedded organization softening, make tubulose, sprawl random alignment by hand, but a plurality of tissue specimens of synchronous detection after the conventional section are mainly used in the character that detects monoclonal antibody.Miler and Groothuis also reported similar method (A.J.C.P.1991,96:228-232), be about to strip of tissue be rolled into cylindric, paraffin embedding, slices across, the section of making a plurality of tissues also can be applicable to the research of antibody.Nineteen ninety Battifora and mithta shred tissue specimen, place respectively among the mould that has groove, add Ago-Gel, investing tissue's sample, and overlapping gel, paraffin is embedding again, makes the section of a plurality of tissues.Sundblad also adopts similar method, and (A.J.C.P.1993,102:192-193), just mould is improved a little.Domestic by retrieval 10 years (1991～2001) patent documentations are not found research report in this respect as yet.
Among the above-mentioned ownership system Preparation Method, the tissue specimen number of placing in a paraffin mass is very limited, tissue dewatering, embedding, fixing method and program are difficult to standardization, the different tissues sample that is in the same section varies in size, spacing differs, global shape is irregular, is difficult to carry out automated analysis.Because the tissue specimen degree of depth in the wax stone differs, different, also be difficult to serial section and carry out solid research (this is crucial for tumor research), and hand skill and labour intensity that need be very high, be difficult to after the random alignment discern specific tissue specimen, detection efficiency is low, and experiment condition is difficult to unified when carrying out a large amount of analyzing specimens, the comparability of experimental result is relatively poor, has limited further developing and using of these technology greatly.And these technology only can be used to detect antibody or be used for the minority immunohistochemistry detect, and are difficult to use in genetic analysiss such as fluorescence in situ hybridization (FISH), original position PCR, original position RT-PCR, original position DNA or mRNA amplification.
Research has for many years confirmed that the generation of tumour and development are rapid, multistage processes of multistep, relate to chromosome and polygenic disappearance, suddenly change, methylate, reset, amplification or inactivation, these variations make the pathway of key signals such as cell growth, apoptosis, differentiation produce obstacle, so far the molecule mechanism of this process complexity is not illustrated as yet fully, and one of its major reason is exactly not set up a kind of method that gene changes in hundreds if not thousands of different tissues samples of analyzing simultaneously.
In tumor research, for the research of the key of adjusting and controlling growth and speed limit step, to the research of the conduction of signal and related gene thereof, to the research of gene structure, quantity, arrangement and variation, all be crucial to the discovery of new gene and the research of adjusting function thereof.These researchs all are based upon on the basis that a large amount of tissue specimens are detected and analyze.Accidental genetic mutation does not often have general significance, and what certain gene appearred in a large amount of different tissue specimens simultaneously unusually then has an important clinic value.Owing to need a large amount of tissue specimens, thereby limited the process of research work.So for the efficient of obtaining, fix, preserving and using that improves tissue specimen, the consumption that reduces sample simultaneously to greatest extent is crucial, the tissue specimen of especially special, rare tumour or other diseases.
Three, technical scheme
At the defective that above-mentioned prior art partly exists, the purpose of this invention is to provide a kind of tissue microarray biochip that is used for multiple biomedical research purpose that adopts the mechanization preparation.The putting in order of dot matrix of organizing of this tissue microarray biochip is predefined, and can carry out multiple assembled arrangement, at least can place different big or small tissue arrays on every tissue microarray biochip, various biological analysis can be carried out, the similarities and differences, variation and mutual relationship thereof and the clinical meaning of the tissue samples of dozens of and even thousands of same type or dissimilar Different Individual can be compared and analyze simultaneously.
To achieve these goals, the technical scheme that the present invention takes is: the tissue microarray biochip that adopts the mechanization preparation, comprise and derive from various malignant tumours, various benign tumours, various precancerous lesions, various non-neoplastic diseases, various normal structures, various embryonic tissues, various blood and hematopoietic tissue, various cell extracts, various biopsies or cell, various hydrothorax, ascites, urine, sputum, cell in the transudate, the histocyte of various in vitro culture and clone, various bacteriums and saccharomycete, the normal structure of various animals, the pathological tissues of various animals, the embryonic tissue of various animals, various animal model tissues, the donor paraffin organization sample of various transgenic animals tissues or various transgenosis organs, be characterized in: adopt mechanization apparatus for preparation or automated preparation instrument from donor paraffin organization sample, to take out cylindric tissue core, each cylindric tissue core is placed in the blank wax stone of another piece (acceptor piece) by design sequence, formative tissue microarray wax stone, from this micro-array tissue wax stone, can obtain a large amount of micro-array tissue sections, section being placed on the slide of silication and gel is the formative tissue micro-array biochip, comprises a large amount of orderly various donor paraffin organization samples on each tissue microarray biochip.
Some other characteristics of the present invention are: donor paraffin organization sample all will be through the privileged site of drawing materials, observation under ethanol or formaldehyde fixed, dehydration, transparent, waxdip, embedding, section, HE dyeing or other specific stains, the optical microscope, diagnosis and position tissue sample will be studied and take a sample.
The area of every kind of micro-array tissue wax stone is respectively 15mm * 15mm, 15mm * 20mm, 15mm * 25mm, 15mm * 30mm, 15mm * 35mm, 15mm * 40mm, 15mm * 45mm, 15mm * 50mm, 20mm * 15mm, 20mm * 20mm, 20mm * 25mm, 20mm * 30mm, 20mm * 35mm, 20mm * 40mm, 20mm * 45mm, 20mm * 50mm; 25mm * 15mm, 25mm * 20mm, 25mm * 25mm, 25mm * 30mm, 25mm * 35mm, 2.5mm * 40mm, 25mm * 45mm, 25mm * 50mm.
The thickness of every kind of micro-array tissue wax stone is respectively 2.0mm, 3.0mm, 4.0mm, 5.0mm, 6.0mm, 7.0mm, 8.0mm, 9.0mm, 10.0mm, 11.0mm, 12.0mm, 13.0mm, 14.0mm, 15.0mm, 16.0mm, 17.0mm, 18.0mm, 19.0mm, 20.0mm.
The cylindric tissue core of being got from the donor paraffin organization sample, its diameter is 0.2mm～4.0mm, that commonly used is 0.5mm～2.0mm; The height of cylindric tissue core is 1.0mm～8.0mm, and often the height that adopts is 2.0mm～5.0mm.
Can place different big or small tissue arrays on every tissue microarray biochip, or place big or small on an equal basis or two big or small tissue arrays of difference, or place three tissue arrays of equal size or different sizes, or place four tissue arrays of equal size or different sizes.
Can take out cylindric tissue core from donor paraffin organization sample, diameter is the cylindric tissue core of 0.2mm, can prepare the micro-array tissue of 2～8000 samples on a slide, and preferable array number is 20～5000 tissue samples; Taking out diameter is the cylindric tissue core of 0.3mm, can prepare the micro-array tissue of 2～5000 samples on a slide, and preferable array number is 20～2180 tissue samples; Taking out diameter is the cylindric tissue core of 0.4mm, can prepare the micro-array tissue of 2～3000 samples on a slide, and preferable array number is 20～1250 tissue samples; Taking out diameter is the cylindric tissue core of 0.5mm, can prepare the micro-array tissue of 2～2000 samples on a slide, and preferable array number is 20～800 tissue samples; Taking out diameter is the cylindric tissue core of 0.6mm, can prepare the micro-array tissue of 2～1500 samples on a slide, and preferable array number is 20～640 tissue samples; Taking out diameter is the cylindric tissue core of 0.7mm, can prepare the micro-array tissue of 2～1200 samples on a slide, and preferable array number is 20～520 tissue samples; Taking out diameter is the cylindric tissue core of 0.8mm, can prepare the micro-array tissue of 2～960 samples on a slide, and preferable array number is 20～450 tissue samples; Taking out diameter is the cylindric tissue core of 0.9mm, can prepare the micro-array tissue of 2～800 samples on a slide, and preferable array number is 20～390 tissue samples; Taking out diameter is the cylindric tissue core of 1.0mm, can prepare the micro-array tissue of 2～640 samples on a slide, and preferable array number is 20～330 tissue samples; Taking out diameter is the cylindric tissue core of 1.1mm, can prepare the micro-array tissue of 2～520 samples on a slide, and preferable array number is 20～300 tissue samples; Taking out diameter is the cylindric tissue core of 1.2mm, can prepare the micro-array tissue of 2～450 samples on a slide, and preferable array number is 20～270 tissue samples; Taking out diameter is the cylindric tissue core of 1.3mm, can prepare the micro-array tissue of 2～390 samples on a slide, and preferable array number is 20～240 tissue samples; Taking out diameter is the cylindric tissue core of 1.4mm, can prepare the micro-array tissue of 2～330 samples on a slide, and preferable array number is 20～210 tissue samples; Taking out diameter is the cylindric tissue core of 1.5mm, can prepare the micro-array tissue of 2～300 samples on a slide, and preferable array number is 20～200 tissue samples; Taking out diameter is the cylindric tissue core of 1.6mm, can prepare the micro-array tissue of 2～270 samples on a slide, and preferable array number is 20～180 tissue samples; Taking out diameter is the cylindric tissue core of 1.7mm, can prepare the micro-array tissue of 2～240 samples on a slide, and preferable array number is 20～160 tissue samples; Taking out diameter is the cylindric tissue core of 1.8mm, can prepare the micro-array tissue of 2～210 samples on a slide, and preferable array number is 20～140 tissue samples; Taking out diameter is the cylindric tissue core of 1.9mm, can prepare the micro-array tissue of 2～200 samples on a slide, and preferable array number is 20～130 tissue samples; Taking out diameter is the cylindric tissue core of 2.0mm, can prepare the micro-array tissue of 2～170 samples on a slide, and preferable array number is 20～125 tissue samples; Taking out diameter is the cylindric tissue core of 2.1mm, can prepare the micro-array tissue of 2～160 samples on a slide, and preferable array number is 20～120 tissue samples; Taking out diameter is the cylindric tissue core of 2.2mm, can prepare the micro-array tissue of 2～140 samples on a slide, and preferable array number is 20～110 tissue samples; Taking out diameter is the cylindric tissue core of 2.3mm, can prepare the micro-array tissue of 2～130 samples on a slide, and preferable array number is 20～105 tissue samples; Taking out diameter is the cylindric tissue core of 2.4mm, can prepare the micro-array tissue of 2～125 samples on a slide, and preferable array number is 20～95 tissue samples; Taking out diameter is the cylindric tissue core of 2.5mm, can prepare the micro-array tissue of 2～120 samples on a slide, and preferable array number is 20～90 tissue samples; Taking out diameter is the cylindric tissue core of 2.6mm, can prepare the micro-array tissue of 2～110 samples on a slide, and preferable array number is 20～85 tissue samples; Taking out diameter is the cylindric tissue core of 2.7mm, can prepare the micro-array tissue of 2～105 samples on a slide, and preferable array number is 20～80 tissue samples; Taking out diameter is the cylindric tissue core of 2.8mm, can prepare the micro-array tissue of 2～95 samples on a slide, and preferable array number is 20～75 tissue samples; Taking out diameter is the cylindric tissue core of 2.9mm, can prepare the micro-array tissue of 2～90 samples on a slide, and preferable array number is 20～70 tissue samples; Taking out diameter is the cylindric tissue core of 3.0mm, can prepare the micro-array tissue of 2～85 samples on a slide, and preferable array number is 20～65 tissue samples; Taking out diameter is the cylindric tissue core of 3.1mm, can prepare the micro-array tissue of 2～80 samples on a slide, and preferable array number is 20～60 tissue samples; Taking out diameter is the cylindric tissue core of 3.2mm, can prepare the micro-array tissue of 2～75 samples on a slide, and preferable array number is 20～55 tissue samples; Taking out diameter is the cylindric tissue core of 3.3mm, can prepare the micro-array tissue of 2～68 samples on a slide, and preferable array number is 20～52 tissue samples; Taking out diameter is the cylindric tissue core of 3.4mm, can prepare the micro-array tissue of 2～65 samples on a slide, and preferable array number is 20～48 tissue samples; Taking out diameter is the cylindric tissue core of 3.5mm, can prepare the micro-array tissue of 2～60 samples on a slide, and preferable array number is 20～44 tissue samples; Taking out diameter is the cylindric tissue core of 3.6mm, can prepare the micro-array tissue of 2～56 samples on a slide, and preferable array number is 20～40 tissue samples; Taking out diameter is the cylindric tissue core of 3.7mm, can prepare the micro-array tissue of 2～52 samples on a slide, and preferable array number is 20～36 tissue samples; Taking out diameter is the cylindric tissue core of 3.8mm, can prepare the micro-array tissue of 2～48 samples on a slide, and preferable array number is 20～32 tissue samples; Taking out diameter is the cylindric tissue core of 3.9mm, can prepare the micro-array tissue of 2～44 samples on a slide, and preferable array number is 20～28 tissue samples; Taking out diameter is the cylindric tissue core of 4.0mm, can prepare the micro-array tissue of 2～40 samples on a slide, and preferable array number is 20～24 tissue samples;
The thickness of every kind of micro-array tissue wax stone is respectively 2.0mm, 3.0mm, 4.0mm, 5.0mm, 6.0mm, 7.0mm, 8.0mm, 9.0mm, 10.0mm, 11.0mm, 12.0mm, 13.0mm, 14.0mm, 15.0mm, 16.0mm, 17.0mm, 18.0mm, 19.0mm, 20.0mm.
The putting in order of dot matrix of organizing of this tissue microarray biochip is predefined, and arranges by following combination: 1). same disease is arranged: be about to be arranged on the tissue microarray biochip with a kind of tissue specimen of Different Individual of disease and the multiple normal control tissue sample of Different Individual; 2). various disease is arranged: soon the tissue specimen of the Different Individual of various disease and the multiple normal control tissue sample of Different Individual are arranged on the tissue microarray biochip; 3). arrange at the same position of same disease: be about to the pathological tissues of same organ, the other tissue of pathology; The multiple normal control tissue sample of normal structure and Different Individual is arranged on the tissue microarray biochip; 4). arrange at the same position of various disease: be about to the pathological tissues of Different Organs, the other tissue of pathology, the multiple normal control tissue sample of normal structure and Different Individual is arranged on the tissue microarray biochip; 5). the same types of organization of same disease arranges: be about to same disease and be arranged on the tissue microarray biochip with the tissue specimen of a kind of types of organization Different Individual and the multiple normal control tissue sample of Different Individual; 6). the same types of organization of various disease arranges: be about to various disease and be arranged on the tissue microarray biochip with the tissue specimen of a kind of types of organization Different Individual and the multiple normal control tissue sample of Different Individual; 7). same disease same clinical stages, arrange: be about to a kind of disease with a kind of clinical stages Different Individual tissue specimen and the multiple normal control tissue sample of Different Individual be arranged on the tissue microarray biochip; 8). various disease same clinical stages, arrange: be about to various disease with a kind of clinical stages Different Individual tissue specimen and the multiple normal control tissue sample of Different Individual be arranged on the tissue microarray biochip; 9). the same sex of same disease is arranged: soon be arranged on the tissue microarray biochip with a kind of tissue specimen of sex Different Individual and the multiple normal control tissue sample of Different Individual with a kind of disease; 10). the same sex of various disease is arranged: be about to various disease and be arranged on the tissue microarray biochip with a kind of tissue specimen of sex Different Individual and the multiple normal control tissue sample of Different Individual; 11). same disease same age bracket is arranged: soon be arranged on the tissue microarray biochip with a kind of tissue specimen of age bracket Different Individual and the multiple normal control tissue sample of Different Individual with a kind of disease; 12). the various disease same age bracket is arranged: be about to various disease and be arranged on the tissue microarray biochip with a kind of tissue specimen of age bracket Different Individual and the multiple normal control tissue sample of Different Individual; 13). arrange before and after the same disease treatment: be about to be arranged on the tissue microarray biochip with the tissue specimen of before a kind of disease treatment or treatment back Different Individual and the multiple normal control tissue sample of Different Individual; 14). arrange before or after the various disease treatment: the tissue specimen of before the various disease treatment soon or treatment back Different Individual and the multiple normal control tissue sample of Different Individual are arranged on the tissue microarray biochip; 15). the same therapeutic modality of same disease is arranged: soon be arranged on the tissue microarray biochip with a kind of tissue specimen of therapeutic modality Different Individual and the multiple normal control tissue sample of Different Individual with a kind of disease; 16). the same therapeutic modality of various disease is arranged: be about to various disease and be arranged on the tissue microarray biochip with a kind of tissue specimen of therapeutic modality Different Individual and the multiple normal control tissue sample of Different Individual; 17). the same molecular labeling of same disease is arranged: be about to contain the tissue specimen of same molecular mark Different Individual and the multiple normal control tissue sample of Different Individual is arranged on the tissue microarray biochip with a kind of disease; 18). the same molecular labeling of various disease is arranged: be about to that various disease contains the tissue specimen of same molecular mark Different Individual and the multiple normal control tissue sample of Different Individual is arranged on the tissue microarray biochip; 19). former of same disease, secondary (transfer), recurrent focus are arranged: be about to be arranged on the tissue microarray biochip with former of a kind of disease Different Individual or the tissue specimen of secondary or recurrent focus and the multiple normal control tissue sample of Different Individual; 20). former of various disease, secondary (transfer), recurrent focus are arranged: be about to former of various disease Different Individual or the tissue specimen of secondary or recurrent focus and the multiple normal control tissue sample of Different Individual and be arranged on the tissue microarray biochip; 21). the same tissue of same disease origin is arranged: be about to organize the tissue specimen of source Different Individual and the multiple normal control tissue sample of Different Individual is arranged on the tissue microarray biochip with a kind of disease with a kind of; 22). the same tissue of various disease origin is arranged: be about to various disease and organize the tissue specimen of source Different Individual and the multiple normal control tissue sample of Different Individual is arranged on the tissue microarray biochip with a kind of; 23). the same differentiation degree of same disease is arranged: soon be arranged on the tissue microarray biochip with a kind of tissue specimen of differentiation degree Different Individual and the multiple normal control tissue sample of Different Individual with a kind of disease; 24). the same differentiation degree of various disease is arranged: be about to various disease and be arranged on the tissue microarray biochip with a kind of tissue specimen of differentiation degree Different Individual and the multiple normal control tissue sample of Different Individual; 25). the same classification of same disease is arranged: soon be arranged on the tissue microarray biochip with the tissue specimen of the same classification Different Individual of a kind of disease and the multiple normal control tissue sample of Different Individual; 26). the same classification of various disease is arranged: be about to the tissue specimen of the same classification Different Individual of various disease and the multiple normal control tissue sample of Different Individual and be arranged on the tissue microarray biochip; 27). same system is arranged: be about to the tissue specimen of various disease Different Individual in the human body same system and the multiple normal control tissue sample of Different Individual and be arranged on the tissue microarray biochip; 28). multisystem is arranged: be about to the tissue specimen of various disease Different Individual in the human body multisystem and the multiple normal control tissue sample of Different Individual and be arranged on the tissue microarray biochip; 29). the arrangement of the identical medical history of same disease: be about to have the tissue specimen of identical medical history Different Individual and the multiple normal control tissue sample of Different Individual and be arranged on the tissue microarray biochip with a kind of disease; 30). the arrangement of the different medical histories of same disease: be about to have the tissue specimen of different medical history Different Individual and the multiple normal control tissue sample of Different Individual and be arranged on the tissue microarray biochip with a kind of disease; 31). the arrangement of the identical infection type of same disease: be about to a kind of disease, have the tissue specimen of Different Individual of identical infection type and the multiple normal control tissue sample of Different Individual and be arranged on the tissue microarray biochip; 32). the arrangement of the different infection types of same disease: be about to be arranged on the tissue microarray biochip with the tissue specimen of the Different Individual of the different infection types of a kind of disease and the multiple normal control tissue sample of Different Individual; 33). same organ normal structure is arranged: the tissue specimen that is about to the same organ Different Individual of human body is arranged on the tissue microarray biochip; 34). the Different Organs normal structure is arranged: the tissue specimen that is about to human body Different Organs Different Individual is arranged on the tissue microarray biochip; 35). embryonic tissue of same stage of development of same organ is arranged: be about to the tissue samples of same embryonic organ's Different Individual of same stage of development and Different Individual adult's multiple normal control tissue sample and be arranged on the tissue microarray biochip; 36). embryonic tissue of same stage of development of Different Organs is arranged: be about to the tissue specimen of same stage of development Different Organs embryonic tissue Different Individual and Different Individual adult's multiple normal control tissue sample and be arranged on the tissue microarray biochip; 37). same organ different developmental phases embryonic tissue is arranged: be about to the tissue specimen of same organ different developmental phases embryonic tissue Different Individual and Different Individual adult's multiple normal control tissue sample and be arranged on the tissue microarray biochip; 38). Different Organs different developmental phases embryonic tissue is arranged: be about to the tissue specimen of Different Organs different developmental phases embryonic tissue Different Individual and the multiple normal adult control tissue sample of Different Individual and be arranged on the tissue microarray biochip; 39). animal tissue arranges: comprising the tissue specimen of the Different Individual of the organs of the various disease of various different animals, the normal structure of Different Organs, the embryonic tissue of different development stage, various transgenic animals and tissue.It arranges basic identical with the arrangement of above-mentioned (1～34) human tissue sample; 40). of the same race with organizing clone to arrange: as to be about to of the same race with organizing the clone sample of in vitro culture to be arranged on the tissue microarray biochip; 41). xenogenesis is with organizing clone to arrange: be about to xenogenesis with organizing the clone sample of in vitro culture to be arranged on the tissue microarray biochip; 42). arrangement mixes in allogenic cell system: be about to be arranged on the tissue microarray biochip with the clone of the various in vitro culture of kind; 43). arrangement mixes in heterogenous cell system: the clone that is about to the various in vitro culture of different genera is arranged on the tissue microarray biochip; 44). microorganism of the same type is arranged: the microbial cell that is about to same type is arranged on the tissue microarray biochip; 45). dissimilar microorganisms are arranged: be about to dissimilar microbial cells and be arranged on the tissue microarray biochip.
The present invention adopts the tissue microarray biochip that is used for multiple biomedical research purpose of mechanization preparation.The putting in order of dot matrix of organizing of this tissue microarray biochip is predefined, and can carry out multiple assembled arrangement, at least can place different big or small tissue arrays on every tissue microarray biochip, can carry out various biological analysis, has a large amount of tissue specimens, can compare and analyze dozens of simultaneously and even thousands ofly carry out various biological analysis these samples, with the mutual relationship and the clinical meaning of the similarities and differences, variation and the disease of same specific molecular or different molecular in the tissue samples of determining Different Individual.
Four, description of drawings
Fig. 1, Fig. 2 are the synoptic diagram of mechanization micro-array tissue preparing instrument;
Fig. 3～Fig. 9 is the work synoptic diagram of mechanization micro-array tissue preparing instrument;
Figure 10 is the synoptic diagram of automatic high-speed micro-array tissue preparing instrument;
Figure 11～Figure 18 is the working state schematic representation of automatic high-speed micro-array tissue preparing instrument;
Figure 19～Figure 42 is that tissue microarray biochip of the present invention is arranged synoptic diagram.
Five, embodiment
The present invention is described in further detail below in conjunction with drawings and Examples.
The needed material of preparation tissue microarray biochip has:
1. the preparation of routine pathology method comprises that the different diseases with various animal all sites of people are planted and the paraffin embedded tissues of different development stage or cell wax stone (being called for short the donor piece).All donor tissue samples that wherein derive from people or other animals are all with corresponding clinical information, comprise age, sex, clinical symptoms, diseased region and pathological diagnosis result comprise tumour or other diseases by stages, classification, differentiation, origin of cell, types of organization and immunohistochemical staining result etc.
2. be used to accept the small cylindric tissue that obtains from the donor piece or the blank wax stone (being called for short the acceptor piece) of cell.
3. be used to diagnose and locate haematine-Yihong (HE) stained that derives from same donor piece.
4. common slide and the silication slide of handling with particular adhesive, corresponding cover glass and mounting glue.
5. the adhesive tape of special processing.
6. come unstuck accordingly and dewaxing agent.
Preparation tissue microarray biochip devices needed comprises:
(1) mechanization micro-array tissue preparing instrument
Referring to Fig. 1, donor piece 1 is placed in the position of container 2, a L shaped guide pole 4 is arranged above the stationary platform 32, position in order to fixed container 2, decompressor 5 is installed on the platform 3, comprise one vertically upward positioning disk 6 and the location-plate 7 on the horizontal direction, location-plate 7 is installed in (not shown) on the X-Y gauge, can carry out the measurement and the adjusting of micron level accurately like this.Vertically-guided dish 6 provides accurate guide surface, and punching press base 8 can slide along flexible track 9 in the plane.9 is the tracks that launch as shown in Figure 2, elastoplast 48 assists punching press base 8 to reset along track 9, impact tube 11 is most advanced and sophisticated sharp keen stainless-steel tube, be placed in above the base 8, impact tube 11 can to dull and stereotyped 3 and container 2 stretch out or withdraw, the endoporus of impact tube 11 is extended down to base 8, and opening 12 is positioned on the lip shape edge 14.
As shown in Figure 1, histotomy through HE dyeing or additive method dyeing can obtain from donor piece 1, and be placed on the slide 13, diagnose and the mark of different colours is carried out at needed diseased region or other positions that will study, represent cancerous tissue, blue colour specification cancer beside organism, yellow expression metastases tissue, the green normal structure or the like of representing as black.The jib 26 that slide 13 is had articulation structure grasps and places (referring to Fig. 9) on the donor 1, clamp 24 on the jib 26 is grasping the edge of transparent microslide 23, jib 54 can change angle in 25 places in the joint, primary importance is fixed in microslide 23 on the container 2, and the second place is removed to appear impact tube 11 microslide 23 horizontal directions.Concrete operations are as follows: container 2 be placed on the platform 3 (referring to Fig. 1) and make its edge near guide pole 4 to guarantee that container 2 is fixed on appointed positions, HE dyeing is carried out in organizing the conventional section of paraffin embedding (tumor sample 15) (3 μ m～5 μ m) and sample 15 is fixed in the donor piece on the slide 13, be positioned over then on the microslide 23, and be positioned (referring to Fig. 9) on the donor piece 1, slide 13 can move on microslide 23, tissue sample 15 in sample 16 and donor piece overlaps fully, as shown in phantom in Figure 9, jib 26 is fixed in primary importance, by the selected diseased region 17 (as when tissue sample is breast cancer; diseased region 17 is exactly the tissue with breast cancer characteristic structure) of pathological diagnosis institute mark, the donor piece 1 below it has the tissue specimen 15 of same structure to be positioned simultaneously.As shown in Figure 1, location-plate 7 can move at the X-Y direction of principal axis under milscale and control lever control.Impact tube 11 is aimed at tissue site selected in the donor piece 1, remove microslide 23, impact tube 11 is squeezed into tissue site (referring to Fig. 2) selected in the donor piece 1, guide bar 6 moves down until dull and stereotyped 8 (preestablishing by installing 5) along track 9, impact tube 11 takes the tissue in the piece of tissue 1 downwards, can obtain tissue sample behind the retraction, stretch into opening 12 with probe and can obtain columniform tissue sample.Put it in shape and the big or small identical acceptor piece 18, acceptor piece 18 can be prepared before sampling, and method of operating is that blank wax stone is placed in the container 2, gets cylindrical hole (as shown in Figure 3) with impact tube 11.
The method that obtains conventional blank matrix wax stone arrangement is: at first impact tube is placed on reference position (as the blank wax stone upper left corner), punch with impact tube, move decompressor or acceptor piece 18 at the X-Y direction of principal axis again, repeat above step, the blank array wax stone (acceptor piece) of different matrixes has just been made.As shown in Figure 3, the aperture is 0.6mm, and the centre distance of Kong Yukong is 0.7mm, and the adjacent holes distance is 0.1mm then.In the ordinary course of things, it is 2mm～6mm highly that cylindrical tissue takes, and radius is 0.3mm～1.0mm.From the marked region sampling of donor piece, and arrange to the new acceptor piece that contains the microarray aperture, its area is 20mm～25mm * 15mm～50mm.The sampling radius can be regulated arbitrarily between 0.1mm～2.0mm, and the radius of taking a sample generally speaking is comparatively suitable between 0.3mm～1.0mm, not only can be unlikely to simultaneously the institutional framework in the former donor piece is caused damage for the usefulness of analysis.On the acceptor piece of 20mm * 45mm, can place the cylindrical tissue specimen more than 1000.
Figure 4 shows that the tissue micro-array wax stone that places sample, before section, earlier the tissue micro-array wax stone is placed on 37 ℃ of incubators 15 minutes, to increase the viscosity between cylindrical tissue samples and the blank wax stone and the pliability of tissue micro-array wax stone, and make its smooth surface smooth, cover with adhesive film 19 again, after adhesive film places, cut the thick micro-array tissue section 20 of 4 μ m～8 μ m along y direction, be positioned on the slide (needing to use the particular adhesive treatment surface) 21, remove adhesive film 19 after smooth, an end 22 of slide is used for identification and label orientation.
(2) automatic high-speed micro-array tissue preparing instrument
Referring to Figure 10, preparing instrument comprises platform 27, contains X gearing 28 and Y gearing 29, has a transmission shaft 30 and 31 separately.Transmission shaft 31 makes to be made platform 32 and moves along Y direction, transmission shaft 30 then makes specimen holder 33 move along X-direction, specimen holder 33 front ends fixing be 3 containers 34,35 and 36, each container is equipped with acceptor paraffin mass 37,38 and 39, donor piece container 40 also is installed in this, donor piece 41 is positioned in the container 40, and contain tissue specimen 42 in the donor piece 41, at its trailing edge is the donor piece carriage 42 and 43 (containing a plurality of containers that hold aqueous sample) of two porous, and a waste canister 44.On dull and stereotyped 27 is a perforating by punching device 45, the latter can move up and down along the z direction of principal axis, perforating by punching device 45 centres are probe carriages 46 of a vertical direction, have a hollow needle 51 on it, perforating by punching device 45 also comprises an inclined-plane acceptor punching frame 48 and an inclined-plane donor punching frame 47, punching frame 48 comprises a mutual lance 49, at its far-end is a tubular acceptor card punch 50, punching frame 47 comprises a mutual lance 53, at its far-end is a tubular donor card punch 52, when mutual lance 49 trails (referring to Figure 11), the opening of tubular acceptor card punch 50 faces hollow needle 51, and when mutual lance 53 stretched, tubular donor card punch 52 was also over against hollow needle 51, and the sequence of operation of perforating device 45 is shown in Figure 11～14.
Figure 10 is mounted package unit, and computer control can make its efficient greatly improve, and can start voluntarily, regulates voluntarily, prepares the micro-array tissue wax stone automatically.The position of specimen holder 33 upper containers among Figure 10 is determined, shown in Figure 11 is the relative position that platform 32 and specimen holder 33 are made in the decision of X-Y regulator, mutual then lance 49 trails, the reference position (1 that tubular acceptor card punch 50 is aimed on the acceptor paraffin mass 37,1), after tubular acceptor card punch 50 puts in place, device will move along the z direction of principal axis, on the acceptor paraffin mass, punch, device 45 is upwards removed acceptor paraffin mass 37, finish punching, X-Y driver adjustment making platform 32 and waste canister 44 are under the tubular acceptor card punch 50 then, the paraffin that probe 46 is released in the hollow needle 51 is examined to waste canister 44, withdrawal hollow needle 51, mutual lance 49 resets, the X-Y driver moves making platform 32 and specimen holder 33 makes donor container 40 place position shown in Figure 13, mutual lance 53 stretches out the tubular card punch 52 of donor in donor paraffin mass 41, perforating device 45 moves down and take cylindrical piece of tissue from donor paraffin mass 41, perforating device 45 is regained the tubular card punch 52 of donor then, the X-Y driver moves makes platform 32 and specimen holder 33 to position shown in Figure 14, punch device 45 moves down, the tubular card punch 52 of donor is placed the corresponding position (1 of donor piece, 1), the tubular card punch 52 of donor is over against hollow needle 51, piece of tissue is dug out tubular card punch 52 of withdrawal donor and mutual lance 53.Said process carries out repeatedly until obtaining enough sample sizes.Make acceptor paraffin mass 37 earlier although be illustrated as, in 38 and 39 one, add tubular tissue again, also can earlier all acceptor paraffin masses be made in advance in fact, add tissue then together, both can reuse piece of tissue 42, also can in donor container 40, add new piece of tissue 41, if the different piece of tissue 41 that more renews after each sampling, each position just includes different tissue samples in the microarray.
Shown in Figure 15 is an orientator, help from sample, to cut selected tissue sample, orientator comprises a microslide 57, between two walls of donor container 40, the tissue sample 42 that comprises skim dyeing above the sample slide 58, use the microscope on 45, can observe and definite tissues needed position, use positioned light source 54 and 55 can determine the correct position of punch device 45 apart from donor paraffin mass 41, when being positioned at desirable position, can focus on a bit from 54 and 55 light beams that send 59 and 61, and then the position of definite mutual lance 53 and 50, it is more satisfactory being placed in the acceptor piece to the cylindrical tissue tight of donor paraffin mass 41, if the tubular card punch 52 of donor has identical interior external diameter with the tubular card punch 50 of acceptor, the size of the tubular tissue of donor is by the decision of card punch internal diameter, and the size in acceptor aperture then by the external diameter decision of card punch, so usually causes receptor hole greater than cylindrical tissue.Therefore the radius of the tubular perforating needle pipe of donor is slightly larger than the radius of the tubular perforating needle pipe of acceptor shown in Figure 16,17, can guarantee that like this donor tubular tissue is identical with the acceptor pore size.After Figure 18 shows that the acceptor wax block filling is organized well, a spot of paraffin is separated tubular tissue, receptor hole slightly is deeper than the donor tubular tissue, between at the bottom of sample and the receptor hole, certain distance is arranged, after micro-array tissue forms, can be from 37 sections of acceptor microarray wax stone, observation sample 42 is determined selected position, also can cut tissue, carry out different molecular biology researches.
Above-mentioned preparation system also comprises computer aided system, with multiple running program, bears different tasks separately, finishes the task that automatic high-speed is made micro-array tissue jointly.
No matter be mechanization or the full-automatic tissue microarray biochip of making, all have identical structure, only be the mode difference of making.The basic structure of tissue microarray biochip comprises following components: the 1. silication slide handled of particular adhesive.2. be stained with the microarray lamellar structure of the 3 μ m～square or rectangular that 8 μ m are thick that downcuts from the microarray wax stone on the silication slide.3. through fixing, come unstuck, the tissue microarray biochip of dewaxing and HE dyeing or other specific stains.Be mainly used in qualitative and diagnosis.4. without coming unstuck dewaxing and HE dyeing, basic structure, arrangement, the identical tissue microarray biochip of the slide of size and content and dyeing.Be mainly used in various biological study purpose.
Tissue microarray biochip be arranged with following mode:
1). same disease is arranged: be about to multiple normal control tissue sample with a kind of tissue specimen of Different Individual of disease and Different Individual and be arranged on the tissue microarray biochip and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is that the difference of Different Individual is by stages or before and after histological types or all ages and classes different sexes or different stage or different therapeutic modality or the treatment or different tissues origin or former, secondary, the cancer of the stomach of recurrence or the tissue specimen of lung cancer or other diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
2). various disease is arranged: soon the multiple normal control tissue sample of the tissue specimen of the Different Individual of various disease and Different Individual is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is the cancer of the stomach of Different Individual or the tissue specimen of other diseases, 30 is the lung cancer of Different Individual or the tissue specimen of other diseases, 30 is the liver cancer of Different Individual or the tissue specimen of other diseases, 30 is the breast cancer of Different Individual or the tissue specimen of other diseases, 30 is the colon cancer of Different Individual or the tissue specimen of other diseases, and 30 is the carcinoma of the rectum of Different Individual or the tissue specimen of other diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
3). arrange at the same position of same disease: the pathological tissues that is about to same organ; the other tissue of pathology, the multiple normal control tissue sample of normal structure and Different Individual is arranged on the tissue microarray biochip (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 60 is the breast cancer of Different Individual or the tissue samples of cancer of the esophagus or other diseases, 60 is the other or other tissue specimen of other pathologies of cancer of same organ, and 60 is the normal structure sample of same organ; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
4). arrange at the same position of various disease: the pathological tissues that is about to Different Organs; the other tissue of pathology, the multiple normal control tissue sample of normal structure and Different Individual is arranged on the tissue microarray biochip (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 20 is the breast cancer of Different Individual or the tissue specimen of other diseases, 20 is the tissue specimen that the breast cancer cancer is other or the other diseases pathology is other of same individuality, and 20 is the breast cancer of same individuality or the normal structure sample of other diseases; 20 is the prostate cancer of Different Individual or the tissue specimen of other diseases, and 20 is the tissue specimen that the prostate cancer cancer is other or the other diseases pathology is other of same individuality, and 20 is the prostate cancer of same individuality or the normal structure sample of other diseases; 20 is the oophoroma of Different Individual or the tissue specimen of other diseases, and 20 is the tissue specimen that the oophoroma cancer is other or the other diseases pathology is other of same individuality, and 20 is the oophoroma of same individuality or the normal structure sample of other diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
5). the same types of organization of same disease arranges: be about to same disease and be arranged on the tissue microarray biochip with the multiple normal control tissue sample of the tissue specimen of a kind of types of organization Different Individual and Different Individual and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is the tissue specimen of lung squamous cancer or adenocarcinoma of lung or other types of organization's diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
6). the same types of organization of various disease arranges: be about to various disease and be arranged on the tissue microarray biochip with the multiple normal control tissue sample of the tissue specimen of a kind of types of organization Different Individual and Different Individual and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is the lung squamous cancer of Different Individual or the tissue specimen of other types disease, 30 is the tissue specimen of the esophagus squama cancer or the other types disease of Different Individual, 30 is the SCC of Different Individual or the tissue specimen of other types disease, 30 is the tissue specimen of the stomach squama cancer or the other types disease of Different Individual, 30 is the tissue specimen of the bile duct squama cancer or the other types disease of Different Individual, and 30 is the tissue specimen of the larynx squama cancer or the other types disease of Different Individual; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
7). same disease same clinical stages, arrange: be about to a kind of disease with a kind of clinical stages Different Individual the tissue specimen and the multiple normal control tissue sample of Different Individual be arranged on the tissue microarray biochip and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is the tissue specimen of cancer of the stomach II phase or cancer of the esophagus III phase or lung cancer IV phase or other diseases same clinical stages; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
8). various disease same clinical stages, arrange: be about to various disease with a kind of clinical stages Different Individual the tissue specimen and the multiple normal control tissue sample of Different Individual be arranged on the tissue microarray biochip and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is the cancer of the stomach II phase or the same tissue specimen by stages of other diseases of Different Individual, 30 is the colon cancer II phase or the same tissue specimen by stages of other diseases of Different Individual, 30 is the lung cancer II phase or the same tissue specimen by stages of other diseases of Different Individual, 30 is the breast cancer II phase or the same tissue specimen by stages of other diseases of Different Individual, 30 is the carcinoma of the rectum II phase or the same tissue specimen by stages of other diseases of Different Individual, and 30 is the prostate cancer II phase or the same tissue specimen by stages of other diseases of Different Individual; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
9). the same sex of same disease is arranged: be about to be arranged on the tissue microarray biochip with the multiple normal control tissue sample of a kind of tissue specimen of sex Different Individual and Different Individual with a kind of disease and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is the tissue specimen of male lung cancer or women's cancer of the stomach or same sex other diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
10). the same sex of various disease is arranged: be about to various disease and be arranged on the tissue microarray biochip with the multiple normal control tissue sample of a kind of tissue specimen of sex Different Individual and Different Individual and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is the male lung cancer of Different Individual or the tissue specimen of same sex other diseases, 30 is the male sex's cancer of the stomach of Different Individual or the tissue specimen of same sex other diseases, 30 is the male sex's liver cancer of Different Individual or the tissue specimen of same sex other diseases, 30 is the male sex's colon cancer of Different Individual or the tissue specimen of same sex other diseases, 30 is the prostate cancer of Different Individual or the tissue specimen of same sex other diseases, and 30 is the male sex's thyroid cancer of Different Individual or the tissue specimen of same sex other diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
11). same disease same age bracket is arranged: be about to be arranged on the tissue microarray biochip with the multiple normal control tissue sample of a kind of tissue specimen of age bracket Different Individual and Different Individual with a kind of disease and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is the tissue specimen of 31 years old～40 years old breast cancer or 41 years old～50 years old prostate cancer or other same diseases of same age bracket; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
12). the various disease same age bracket is arranged: be about to various disease and be arranged on the tissue microarray biochip with the multiple normal control tissue sample of a kind of tissue specimen of age bracket Different Individual and Different Individual and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is Different Individual 31 years old～40 years old the breast cancer or the tissue specimen of same age bracket other diseases, 30 is Different Individual 31 years old～40 years old the prostate cancer or the tissue specimen of same age bracket other diseases, 30 is Different Individual 31 years old～40 years old the lung cancer or the tissue specimen of same age bracket other diseases, 30 is Different Individual 31 years old～40 years old the cancer of the stomach or the tissue specimen of same age bracket other diseases, 30 is Different Individual 31 years old～40 years old the colon cancer or the tissue specimen of same age bracket other diseases, and 30 is Different Individual 31 years old～40 years old the liver cancer or the tissue specimen of same age bracket other diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
13). arrange before and after the same disease treatment: be about to multiple normal control tissue sample with the tissue specimen of before a kind of disease treatment or treatment back Different Individual and Different Individual and be arranged on the tissue microarray biochip and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is before the liver cancer treatment or after the bladder cancer treatment or before other same disease treatments or the tissue specimen after the treatment; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
14). arrange before or after the various disease treatment: the tissue specimen of before the various disease treatment soon or treatment back Different Individual and the multiple normal control tissue sample of Different Individual are arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 be before the liver cancer treatment of Different Individual or treatment back or other diseases treatment before or the tissue specimen after the treatment, 30 be before the bladder cancer treatment of Different Individual or treatment back or other diseases treatment before or the tissue specimen after the treatment, 30 is before the oophoroma treatment of Different Individual or before treatment back or the other diseases treatment or the tissue specimen after the treatment, 30 be before the treatment of pancreatic cancer of Different Individual or treatment back or other diseases treatment before or the tissue specimen after the treatment, 30 be before the lung cancer therapy of Different Individual or treatment back or other diseases treatment before or the tissue specimen after the treatment, 30 is before the prostate cancer therapy of Different Individual or before treatment back or the other diseases treatment or the tissue specimen after the treatment; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
15). the same therapeutic modality of same disease is arranged: be about to be arranged on the tissue microarray biochip with the multiple normal control tissue sample of a kind of tissue specimen of therapeutic modality Different Individual and Different Individual with a kind of disease and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is that carcinoma of testis chemotherapy or kidney radiation therapy or other diseases adopt the tissue specimen after same mode is treated; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
16). the same therapeutic modality of various disease is arranged: be about to various disease and be arranged on the tissue microarray biochip with the multiple normal control tissue sample of a kind of tissue specimen of therapeutic modality Different Individual and Different Individual and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is that the carcinoma of testis chemotherapy of Different Individual or other diseases adopt the tissue specimen after same mode is treated, 30 is that the kidney chemotherapy of Different Individual or other diseases adopt the tissue specimen after same mode is treated, 30 is that the thyroid cancer chemotherapy of Different Individual or other diseases adopt the tissue specimen after same mode is treated, 30 is that the liver cancer chemotherapy of Different Individual or other diseases adopt the tissue specimen after same mode is treated, 30 is that the oophoroma chemotherapy of Different Individual or other diseases adopt the tissue specimen after same mode is treated, and 30 is that the breast cancer chemotherapy of Different Individual or other diseases adopt the tissue specimen after same mode is treated; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
17). the same molecular labeling of same disease is arranged: be about to be arranged on the tissue microarray biochip with the multiple normal control tissue sample that a kind of disease contains the tissue specimen of same molecular mark Different Individual and Different Individual and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is the tissue specimen of the colon cancer CEA positive or the cancer of the stomach ErbB-2 positive or the same molecular labeling positive of other diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
18). the same molecular labeling of various disease is arranged: be about to multiple normal control tissue sample that various disease contains the tissue specimen of same molecular mark Different Individual and Different Individual and be arranged on the tissue microarray biochip and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is the tissue specimen of the colon cancer CEA positive or the same molecular labeling positive of other diseases of Different Individual, 30 is the tissue specimen of the carcinoma of the rectum CEA positive or the same molecular labeling positive of other diseases of Different Individual, 30 is the tissue specimen of the cervical carcinoma CEA positive or the same molecular labeling positive of other diseases of Different Individual, 30 is the tissue specimen of the cancer of the stomach CEA positive or the same molecular labeling positive of other diseases of Different Individual, 30 is the tissue specimen of the liver cancer CEA positive or the same molecular labeling positive of other diseases of Different Individual, and 30 is the tissue specimen of the nasopharyngeal carcinoma CEA positive or the same molecular labeling positive of other diseases of Different Individual; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
19). former of same disease, secondary (transfer), recurrent focus are arranged: be about to multiple normal control tissue sample with the tissue specimen of former of a kind of disease Different Individual or secondary or recurrent focus and Different Individual and be arranged on the tissue microarray biochip and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is the tissue specimen of liver cancer primary lesion or lung cancer metastasis focus or breast cancer relapse focus; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
20). former of various disease, secondary (transfer), recurrent focus are arranged: the multiple normal control tissue sample that is about to the tissue specimen of former of the various disease Different Individual or secondary or recurrent focus and Different Individual is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is former of liver cancer or the focus of secondary or recurrence or former or the secondary or the recurrent focus tissue of other diseases of Different Individual; 30 is former of the lung cancer of Different Individual or former or secondary or recurrent focus tissue of secondary or recurrent focus or other diseases; 30 is former of the breast cancer of Different Individual or former or secondary or recurrent focus tissue of secondary or recurrent focus or other diseases; 30 is former of the colon cancer of Different Individual or former or secondary or recurrent focus tissue of secondary or recurrent focus or other diseases; 30 is former of the thyroid cancer of Different Individual or former or secondary or recurrent focus tissue of secondary or recurrent focus or other diseases; 30 is former of the oophoroma of Different Individual or former or secondary or recurrent focus tissue of secondary or recurrent focus or other diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
21). the same tissue of same disease origin is arranged: be about to be arranged on the tissue microarray biochip with the multiple normal control tissue sample of a kind of tissue specimen of organizing the source Different Individual and Different Individual with a kind of disease and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is to originate from the lung cancer of epithelial tissue or originate from the leiomyosarcoma of mesenchymal tissue or the tissue specimen of the other diseases of same tissue origin; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
22). the same tissue of various disease origin is arranged: be about to various disease and be arranged on the tissue microarray biochip with the multiple normal control tissue sample of a kind of tissue specimen of organizing the source Different Individual and Different Individual and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is the tissue specimen that Different Individual originates from the lung cancer or the same origin of other diseases of epithelial tissue, 30 is that Different Individual originates from the cholangiocarcinoma of epithelial tissue or the tissue specimen of the same origin of other diseases, 30 is the tissue specimen that Different Individual originates from the liver cancer or the same origin of other diseases of epithelial tissue, 30 is the tissue specimen that Different Individual originates from the cervical carcinoma or the same origin of other diseases of epithelial tissue, 30 is the tissue specimen that Different Individual originates from the thyroid cancer or the same origin of other diseases of epithelial tissue, and 30 is the tissue specimen that Different Individual originates from the colon cancer or the same origin of other diseases of epithelial tissue; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
23). the same differentiation degree of same disease is arranged: be about to be arranged on the tissue microarray biochip with the multiple normal control tissue sample of a kind of tissue specimen of differentiation degree Different Individual and Different Individual with a kind of disease and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is high differentiation lung squamous cancer or low differentiation sdenocarcinoma of stomach or with a kind of tissue specimen of other differentiation types of disease; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
24). the same differentiation degree of various disease is arranged: be about to various disease and be arranged on the tissue microarray biochip with the multiple normal control tissue sample of a kind of tissue specimen of differentiation degree Different Individual and Different Individual and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is the height differentiation lung squamous cancer of Different Individual or the tissue specimen of the same differentiation type of other diseases, 30 is the height differentiation liver cancer of Different Individual or the tissue specimen of the same differentiation type of other diseases, 30 is the height differentiation breast cancer of Different Individual or the tissue specimen of the same differentiation type of other diseases, 30 is the height differentiation cancer of pancreas of Different Individual or the tissue specimen of the same differentiation type of other diseases, 30 is the tissue specimen of the high score cancer of the esophagus or the same differentiation type of other diseases of Different Individual, and 30 is the height differentiation oophoroma of Different Individual or the tissue specimen of the same differentiation type of other diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
25). the same classification of same disease is arranged: be about to multiple normal control tissue sample with the tissue specimen of the same classification Different Individual of a kind of disease and Different Individual and be arranged on the tissue microarray biochip and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is the tissue specimen of adenocarcinoma of lung II level or stomach squama cancer III level or the same classification of other diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
26). the same classification of various disease is arranged: soon the multiple normal control tissue sample of the tissue specimen of the same classification Different Individual of various disease and Different Individual is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is the adenocarcinoma of lung II level of Different Individual or the tissue specimen of the same classification of other diseases, 30 is the esophagus squama cancer II level of Different Individual or the tissue specimen of the same classification of other diseases, 30 is the liver cancer II level of Different Individual or the tissue specimen of the same classification of other diseases, 30 is the cancer of pancreas II level of Different Individual or the tissue specimen of the same classification of other diseases, 30 is the thyroid cancer II level of Different Individual or the tissue specimen of the same classification of other diseases, and 30 is the tissue specimen of Different Individual breast cancer II level or the same classification of other diseases; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
27). same system is arranged: soon the multiple normal control tissue sample of the tissue specimen of the interior various disease Different Individual of human body same system and Different Individual is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 220 different samples, and wherein 50 is cancer of the esophagus, and 50 is cancer of the stomach, and 50 is colon cancer, and 50 is rectum cancer tissue's sample; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
28). multisystem is arranged: soon the multiple normal control tissue sample of the tissue specimen of the interior various disease Different Individual of human body multisystem and Different Individual is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 220 different samples, wherein 50 is the tissue specimen of the digestive system cancer of the esophagus or the other system disease of Different Individual, 50 is the tissue specimen of the respiratory system lung cancer or the other system disease of Different Individual, 50 is the tissue specimen of the urinary system kidney or the other system disease of Different Individual, and 50 is the tissue specimen of the central nervous system retinoblastoma or the other system disease of Different Individual; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
29). the arrangement of the identical medical history of same disease: be about to a kind of disease, the multiple normal control tissue sample with the tissue specimen of identical medical history Different Individual and Different Individual is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is to have the lung cancer of Different Individual of 5 years medical histories of smoking or the tissue specimen that other diseases has identical medical history; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
30). the arrangement of the different medical histories of same disease: be about to a kind of disease, the multiple normal control tissue sample with the tissue specimen of different medical history Different Individual and Different Individual is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 220 different samples, wherein 50 is the cancerous lung tissue sample of the Different Individual in 5 years of smoking, 50 is the cancerous lung tissue sample of the Different Individual in 10 years of smoking, 50 is the cancerous lung tissue sample of the Different Individual in 15 years of smoking, and 50 is the cancerous lung tissue sample of the vicennial Different Individual of smoking; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
31). the arrangement of the identical infection type of same disease: be about to a kind of disease, the multiple normal control tissue sample with the tissue specimen of Different Individual of identical infection type and Different Individual is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 be hepatitis B virus positive Different Individual the liver cancer tissue sample or infect the tissue specimen of other viral Different Individual; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
32). the arrangement of the different infection types of same disease: be about to multiple normal control tissue sample with the tissue specimen of the Different Individual of the different infection types of a kind of disease and Different Individual and be arranged on the tissue microarray biochip and (also can not contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 60 is the liver cancer tissue sample of the Different Individual of hepatitis B virus positive, 60 is the liver cancer tissue sample of the Different Individual of the hepatitis C virus positive, and 60 is the liver cancer tissue sample of the Different Individual of the fourth hepatovirus positive; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
33). same organ normal structure is arranged: soon the multiple normal control tissue sample of the tissue specimen of the same organ Different Individual of human body and Different Individual is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is normal liver or normal lungs or other organs and tissues samples of Different Individual; 20 is the normal structure that the normal structure check sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
34). the Different Organs normal structure is arranged: soon the multiple normal control tissue sample of the tissue specimen of human body Different Organs Different Individual and Different Individual is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is liver or other normal structure samples of Different Individual, 30 is spleen or other normal structure samples of Different Individual, 30 is lungs or other normal structure samples of Different Individual, 30 is pancreas or other normal structure samples of Different Individual, 30 is ovary or other normal structure samples of Different Individual, and 30 is prostate or other normal structure samples of Different Individual; 20 is the normal structure that the normal structure contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
35). embryonic tissue of same stage of development of same organ is arranged: the multiple normal control tissue sample that is about to the tissue samples of same embryonic organ's Different Individual of same stage of development and Different Individual adult is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 180 is 3 months fetal livers or 5 months embryo's lungs or the embryonic tissue sample of other same stages of development of internal organs; 20 normal structure contrast samples that are Different Individual is grown up comprise the normal structure at the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
36). embryonic tissue of same stage of development of Different Organs is arranged: the multiple normal control tissue sample that is about to the tissue specimen of same stage of development Different Organs embryonic tissue Different Individual and Different Individual adult is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is 3 months fetal livers or the embryonic tissue sample of other same stages of development of organ of Different Individual, 30 is 3 months embryo's lungs or the embryonic tissue sample of other same stages of development of organ of Different Individual, 30 is 3 months embryo's spleens or the embryonic tissue sample of other same stages of development of organ of Different Individual, 30 is 3 months embryo and brain tissues or the embryonic tissue sample of other same stages of development of organ of Different Individual, 30 is 3 months embryo's kidneys or the embryonic tissue sample of other same stages of development of organ of Different Individual, and 30 is 3 months embryo hearts or the embryonic tissue sample of other same stages of development of organ of Different Individual; 20 normal structure contrast samples that are Different Individual is grown up comprise the normal structure at the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
37). same organ different developmental phases embryonic tissue is arranged: the multiple normal control tissue sample that is about to the tissue specimen of same organ different developmental phases embryonic tissue Different Individual and Different Individual adult is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 220 different samples, and wherein 50 is 3 months fetal livers, and 50 is 5 months fetal livers, and 50 is 7 months fetal livers, and 50 is the tissue specimen of 9 months fetal livers or other internal organs; 20 normal structure contrast samples that are Different Individual is grown up comprise the normal structure at the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
38). Different Organs different developmental phases embryonic tissue is arranged: soon the multiple normal adult control tissue sample of the tissue specimen of Different Organs different developmental phases embryonic tissue Different Individual and Different Individual is arranged on the tissue microarray biochip and (also can contain the normal control tissue sample).As contain the tissue microarray biochip of 200 different samples, wherein 30 is 3 months fetal livers of Different Individual, 30 is 4 months embryo's spleens, 30 is 5 months embryo's lungs, 30 is 6 months embryo's kidneys of Different Individual, 30 is 7 months fetal pancreas, 30 is 8 months embryo hearts, and 20 is the normal structure that the normal adult tissue contrast sample of Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, pancreas or other positions.
39). animal tissue arranges: comprising the tissue specimen of the Different Individual of the organs of the various disease of various different animals, the normal structure of Different Organs, the embryonic tissue of different development stage, various transgenic animals and tissue.It arranges basic identical with the arrangement of above-mentioned (1～34) human tissue sample.
40). of the same race with organizing clone to arrange: as to be about to of the same race with organizing the clone sample of in vitro culture to be arranged on the tissue microarray biochip.As contain the tissue microarray biochip of 50 different samples, wherein 40 is human hepatoma cell strain or stomach cancer cell line or other tissue-derived cell lines of separate sources; 10 different non-aforementioned tissue-derived cell lines in contrast.
41). xenogenesis is with organizing clone to arrange: be about to xenogenesis with organizing the clone sample of in vitro culture to be arranged on the tissue microarray biochip.As contain the tissue microarray biochip of 50 different samples, and wherein 20 is the human hepatoma cell strain of separate sources, 20 is the rat liver cancer cell line of separate sources or the tumor cell line in other kinds source; 10 different non-aforementioned tissue-derived cell lines in contrast.
42). arrangement mixes in allogenic cell system: be about to be arranged on the tissue microarray biochip with the clone of the various in vitro culture of kind.As contain the tissue microarray biochip of 50 different samples, and wherein 10 is the Bel7402, and 10 is the SGC-7901, and 10 is MCF-7, and 10 is human cervical carcinoma cell system; Other people source cell in 10 different non-above-mentioned sources is in contrast.
43). arrangement mixes in heterogenous cell system: the clone that is about to the various in vitro culture of different genera is arranged on the tissue microarray biochip.As contain the tissue microarray biochip of 50 different samples, and wherein 10 is the Bel7402, and 10 is the mouse hepatoma cell line, and 10 is the rabbit hepatoma cell line, and 10 is the dog hepatoma cell line; Other people source cell in 10 different non-above-mentioned sources is in contrast.
44). microorganism of the same type is arranged: the microbial cell that is about to same type is arranged on the tissue microarray biochip.As contain the tissue microarray biochip of 50 different samples, wherein 40 is various different coccuses or bacillus or other microbial cell samples; 10 different non-aforementioned micro organism cells in contrast.
45). dissimilar microorganisms are arranged: be about to dissimilar microbial cells and be arranged on the tissue microarray biochip.As contain the tissue microarray biochip of 50 different samples, and wherein 10 is various different coccuses, and 10 is bacillus, and 10 is spirillum, and 10 is Chlamydia or other microorganisms; 10 different non-aforementioned micro organism cells in contrast.
The area size of micro-array tissue wax stone is respectively: 15mm * 15mm, 15mm * 20mm, 15mm * 25mm, 15mm * 30mm, 15mm * 35mm, 15mm * 40mm, 15mm * 45mm, 15mm * 50mm, 20cm * 15mm, 20mm * 20cm, 20cm * 25cm, 20cm * 30cm, 20cm * 35cm, 20cm * 40cm, 20cm * 45cm, 20cm * 50cm; 25cm * 15mm, 25mm * 25cm, 25cm * 30cm, 25cm * 35cm, 25cm * 40cm, 25cmm * 45cm, 25cm * 50cm.
The thickness of micro-array tissue wax stone is respectively 2.0mm, 3.0mm, 4.0mm, 5.0mm, 6.0mm, 7.0mm, 8.0mm, 9.0mm, 10.0mm, 11.0mm, 12.0mm, 13.0mm, 14.0mm, 15.0mm, 16.0mm, 17.0mm, 18.0mm, 19.0mm, 20.0mm.
The size of dot matrix is relevant with the internal diameter of sampling probe on the tissue microarray biochip, arranges as follows:
Minimum and the best point sample number of maximum point sample number (individual) (individual) of sampling needle inside radius (mm)
0.10?????????????????????2～8000???????????????????????≤5000
0.15?????????????????????2～5000???????????????????????≤2180
0.20?????????????????????2～3000???????????????????????≤1250
0.25?????????????????????2～2000???????????????????????≤800
0.30?????????????????????2～1500???????????????????????≤640
0.35????????????????????2～1200???????????????????????≤520
0.40????????????????????2～960????????????????????????≤450
0.45????????????????????2～800????????????????????????≤390
0.50????????????????????2～640????????????????????????≤330
0.55????????????????????2～520????????????????????????≤300
0.60????????????????????2～450????????????????????????≤270
0.65????????????????????2～390????????????????????????≤240
0.70????????????????????2～330????????????????????????≤210
0.75????????????????????2～300????????????????????????≤200
0.80????????????????????2～270????????????????????????≤180
0.85????????????????????2～240????????????????????????≤160
0.90????????????????????2～210????????????????????????≤136
0.95????????????????????2～200????????????????????????≤130
1.00????????????????????2～170????????????????????????≤128
1.05????????????????????2～160????????????????????????≤118
1.10????????????????????2～136????????????????????????≤108
1.15????????????????????2～130????????????????????????≤102
1.20????????????????????2～128????????????????????????≤94
1.25????????????????????2～118????????????????????????≤88
1.30????????????????????2～108????????????????????????≤82
1.35????????????????????2～102????????????????????????≤78
1.40????????????????????2～94?????????????????????????≤72
1.45????????????????????2～88?????????????????????????≤68
1.50????????????????????2～82?????????????????????????≤64
1.55????????????????????2～78?????????????????????????≤60
1.60????????????????????2～72?????????????????????????≤56
1.65????????????????????2～68?????????????????????????≤52
1.70????????????????????2～64?????????????????????????≤48
1.75????????????????????2～60?????????????????????????≤44
1.80????????????????????2～56?????????????????????????≤40
1.85????????????????????2～52?????????????????????????≤36
1.90????????????????????2～48?????????????????????????≤32
1.95????????????????????2～44?????????????????????????≤28
2.00????????????????????2～40?????????????????????????≤24
The thickness of tissue microarray biochip section is generally 2 μ m～8 μ m, 3 μ m commonly used～6 μ m, the most frequently used 5 μ m.The distribution of tissue microarray biochip mainly comprises: a part is arranged: promptly contain a part of the different equidistant arrangement of dot matrix number on a tissue microarray biochip, as Figure 19, Figure 20 and Figure 21; Two parts are arranged: contain on a tissue microarray biochip promptly that the dot matrix number equates or two parts of unequal equidistant arrangement, as Figure 22～Figure 27: three parts are arranged: contain on a tissue microarray biochip promptly that the dot matrix number equates or three parts of unequal equidistant arrangement, as Figure 28～Figure 31; Four parts are arranged: contain on a tissue microarray biochip promptly that the dot matrix number equates or four parts of unequal equidistant arrangement, as Figure 32～Figure 42.
Tissue microarray biochip provides a kind of high power capacity, and the tissue specimen of large sample can carry out correlation analysis to a large amount of specific tissue specimens simultaneously.Tissue microarray biochip can be for the normal person, the growth period that the normal person is different, the stage of development that the embryo is different, the tumour of various animals, the other diseases of various animals, the growth period that animal is different, the application of the different sheets of animal embryo not only is confined to the research of human tumor, also is applicable to the stage of development of human other diseases, various transgenic animals, the various tissues or the clone of in vitro culture, various bacteriums, virus, the activation of the range gene of fungi and other microorganisms, disappearance, sudden change, various mRNA express, various protein moleculars qualitative, quantitatively, the location and with the correlationship research of cell function.
In addition, tissue microarray biochip can be used for a large amount of genomes (as dna sequencing) analysis and the analysis of specific gene target spot.Can be used for assessing tumour diagnostic reagent or other diseases diagnostic reagent, as specific antibody or special genes probe.Tissue microarray biochip can carry out one or more DNA, RNA or protein analysis in dozens of and even the thousands of known cancer tissue simultaneously, all is very useful for the diagnosis index of determining tumour or other diseases, prognostic indicator or treatment prediction index.Tissue microarray biochip also helps to formulate the tumour patient with specific molecule markers or the personalized therapy program of other diseases.Also can be used for seeking the target spot of new gene therapy and design different therapeutic schemes with this.
Tissue microarray biochip can carry out various biological analysis, comprising:
(1) can carry out HE dyeing
The grown form of main display organization cell and the basic structure of nucleus, tenuigenin and cytoplasm.Can be used for diagnosis, antidiastole, judge the histological type of tissue and cell, judge the origin and the differentiation of tissue, can compare and analyze the similarities and differences, variation and mutual relationship thereof and the clinical meaning of the tissue samples of dozens of and even thousands of same type or dissimilar Different Individual simultaneously.
(2) can carry out various specific stains
Main demonstration pathogenic microorganism and metabolic product, protozoon and parasite, endogenous pigment, artificial pigementation particle, dna exogenous pigment and inorganic salts, show muscle fibre, collagenous fibres, reticulum fiber and elastic fibers, amyloid, carbohydrates, neutral fat and lipid, protein and nucleic acid and the material that contains the phenolic group group.Can be used for diagnosing various pathogeny microorganisms, protozoon and parasitic infection disease, particular matter precipitation or accumulation property disease are distinguished epithelium or non-epithelial tumour, and diagnosis contains the tumour of special construction and material etc.Can compare simultaneously and analyze dozens of and even thousands of same type or dissimilar Different Individual or the similarities and differences, variation and mutual relationship thereof and the clinical meaning of the interior exotic matter of different tissue samples developing period.
(3) can carry out various enzyme stainings
Main activity or the content that shows various redox enzymes, group's transferase, hydrolase and synthetic enzyme.Be usually used in the diagnosis of enzymatic activity abnormality disease.Can compare simultaneously with the similarities and differences of analyzing dozens of and even thousands of same type or dissimilar Different Individual or the interior enzymatic activity of different tissue samples developing period, variation and with the mutual relationship and the clinical meaning of disease.
(4) can carry out immunohistochemical staining
Nearly all albumen and polypeptide class antigen molecule in the main showed cell nuclear, on the nuclear membrane, in the tenuigenin, on the cell membrane, in the organelle, on the organelle coating and in the tissue.The mark that mainly comprises epithelial tissue and related neoplasms thereof, the label of soft tissue and related neoplasms thereof, lymphohematopoietic tissue delivered and relevant tumor marker thereof, the antigenic mark of neuroendocrine cell and acceptor thereof, various cell factors and associated receptor thereof, various enzymes and human tissue compatibility antigen and various pathogenic microorganism comprise the specific antigen of bacterium, virus, mould, protozoon, parasite etc.Be usually used in diagnosis, the antidiastole of tumour and non-neoplastic disease, the selection and the prognosis evaluation of therapeutic modality, judge the origin and the differentiation of cell, determine the tissue and the interior location of cell of various related antigen molecules, study the correlationship of cellular morphology and function, study the correlationship of various protein moleculars and cell growth, growth, metabolism, differentiation; Whether the content of studying various antigen molecules increases, minimizing or whether the site changes and with the correlationship of cell function and disease.What is more important can compare simultaneously with the tissue samples of analyzing dozens of and even thousands of same type or dissimilar Different Individual or the Different Individual of the Different Individual of same developing period or different developing periods in same specific molecular or different molecular the similarities and differences, variation and with the mutual relationship and the clinical value of disease.
(5) can carry out nucleic acid hybridization in situ detects
In situ hybridization comprises DNA-DNA, DNA-RNA, RNA-RNA hybridization.Its concrete grammar comprises fluorescence in situ hybridization (FISH), the original position DNA synthetic (PRINS) that original position PCR, original position RT-PCR, oligonucleotides start etc.Main demonstration chromosome, a certain zone of chromosome, the change on chromosome structure and the number; Show genome on the chromosome, gene, the change on quantity, structure or function of genetic fragment or a certain site.Show the change that gene expression is normal, cross expression, expression deletion or expression.Be usually used in detecting the expression analysis of various oncogenes of tumor tissues and antioncogene; Special genes is to the influence of tissue development in the detection embryonic tissue; Study the mechanism of action of nucleotide sequence in some tumour or non-neoplastic disease of special pathogenic microorganism; The expression of inquiring into various endocrine hormones and various cytokine gene and corresponding acceptor gene reaches nerve, endocrine, hematopoiesis and immune adjusting and influence; The index and the medicine of screening and evaluation diagnosing tumour disease; The potential target of screening and evaluation prediction and treatment various diseases; Estimate and select the therapeutic scheme of various diseases; The index of screening and evaluation treatment monitoring and prognosis evaluation.Simultaneously parallel analysis dozens of and even thousands of same type or dissimilar Different Individual, the similarities and differences that same specific gene or different genes are expressed in the tissue specimen of same type or dissimilar same or different developing periods, variation, level and with the mutual relationship and the clinical meaning of disease.Hybridization in situ technique also can with immunohistochemistry technique, DNA chip technology, cDNA array technique, the mRNA dot matrix techniques, PCP skill sample, Southern blotting, technology such as Northern blotting and Westernblotting are used in combination, and can obviously improve the efficient of detection, improve the susceptibility and the accuracy that detect, help lend some impetus to the process of research.
The inventor has provided following embodiment, but the invention is not restricted to these embodiment.
The HE dyeing comparative diagnoses of embodiment 1. tissue microarray biochips
For different tumor tissues or other tissue specimens are further diagnosed and are compared simultaneously and analyze, all tissue specimens are all through ethanol or formaldehyde fixed, paraffin embedding, HE dyeing diagnosis location, be prepared into the multisample tumor tissues micro-array biochip that the sample diameter is the Different Individual of 1.5mm with mechanization or robotization array preparing instrument then, the stomach organization sample that contains 180 Different Individual on the chip, 40 routine papillary adenocarcinomas wherein, 31 routine tubular adenocarcinomas, 25 routine myxoadenocarcinomas, 16 routine signet ring cell cancers, the low partization gland cancer of 15 examples, 14 routine undifferentiated carcinomas, 14 routine adenosquamous carcinomas, 13 routine squama type cell cancers, 12 routine carcinoids; The normal structure contrast sample of 20 Different Individual comprises the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, the normal structure at pancreas or other positions also can prepare the tissue microarray biochip that other types are arranged, and carries out HE dyeing according to conventional method.At first tissue microarray biochip is dewaxed, dehydration, transparent, aquation, soak 5-10 minute after washing with the haematine dye liquor, use hydrochloride alcohol (70% alcohol 99ml, concentrated hydrochloric acid 1ml) the differentiation several seconds then, washing.Return blue a moment (also can in tap water, return blue 5 minutes) with 1% ammoniacal liquor.After returning basket, whether the dyeing of test under microscope nucleus is appropriate.If hyperchromasia need repeat differentiation; If cross light, but dual-staining then.And then wash, soaked 2 minutes washing, dehydration, transparent, sealing at last with eosin stain.The HE of tissue microarray biochip the dye grown form and the nucleus of main display organization and cell, the basic structure of tenuigenin and cytoplasm.Can be used for diagnosis, antidiastole, judge the histological type of tissue and cell, judge the origin and the differentiation of tissue, can compare simultaneously and the similarities and differences of the tissue specimen of analyzing dozens of and even thousands of same type or dissimilar and various different arrangements, variation, and the mutual relationship and the clinical meaning of disease.
The specific stain comparative diagnoses of embodiment 2. tissue microarray biochips
For some special structure and material in the tumor tissues of Different Individual or other histocytes are further diagnosed and compared simultaneously and analyze, must take special colouring method to show.All tissue specimens are all through ethanol or formaldehyde fixed, paraffin embedding, HE dyeing diagnosis location is prepared into multisample tumor tissues or other tissue microarray biochips that the sample diameter is the Different Individual of 1.3mm with mechanization or robotization array preparing instrument then, contains the leiomyosarcoma tissue sample of 180 Different Individual on the chip, 20 is the normal structure contrast sample of Different Individual, comprise the heart, brain, muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, the normal structure at pancreas or other positions, also can prepare the tissue microarray biochip that other types are arranged, carry out muscle fibre dyeing according to conventional method.At first tissue microarray biochip is dewaxed, dehydration, transparent, aquation, after handling 30 minutes (can save this step) with acid potassium bichromate solution through the liquid-solid fixed tissue of Zenker, washed from the beginning 2 minutes, use 1 minute after washing of acid potassium permanganate aqueous solution chlorination again, by 1% oxalic acid bleaching row washing after 1 minute, spend the night 95% alcohol and anhydrous alcohol dehydration with the dip-dye of phosphotungstic acid haematoxylin, dimethylbenzene is transparent, the neutral gum sealing.Can show leiomyosarcoma muscle fibre situation arranged evenly.But if adopt also pathogenic microorganism and metabolic product, protozoon and the parasite in the display organization cell of other specific stains, endogenous pigment, artificial pigementation particle, dna exogenous pigment and inorganic salts, show muscle fibre, collagenous fibres, reticulum fiber, elastic fibers and amyloid, carbohydrates, neutral fat and lipid, protein and nucleic acid and the material that contains the phenolic group group.The specific stain of tissue microarray biochip can be used for diagnosing various pathogenic microorganisms, protozoon and parasitic infection disease, and particular matter precipitation or accumulation property disease are distinguished epithelium or non-epithelial tumour, and diagnosis contains the tumour of special construction and material etc.Can compare simultaneously with analyze dozens of and even thousands of same type or dissimilar Different Individual or different developing period or other various different tissue samples of arranging in the similarities and differences, variation, and the mutual relationship and the clinical meaning of disease of exotic matter.
Embodiment 3: enzyme staining's comparative diagnoses of tissue microarray biochip
For the activity of various enzymes in different tumor tissues or other histocytes is measured,, must adopt enzyme histochemistry's method to dye so that further diagnose and simultaneously a large amount of tissue specimens is compared and analyzes.All tissue specimens are all fixed through ethanol, adopt mechanization or robotization array preparing instrument to be prepared into the multisample tumor tissues micro-array biochip that the sample diameter is the Different Individual of 1.2mm then, the various tumor tissues samples that contain 180 Different Individual on the chip, wherein 50 is that the liver cancer tissue sample of Different Individual (comprises 43 hepatocellular carcinomas, 5 cholangiocellular carcinomas and 2 cell mixing cancers), 50 is that the stomach organization sample of Different Individual (comprises 21 routine papillary adenocarcinomas, 9 routine tubular adenocarcinomas, 7 routine myxoadenocarcinomas, 5 routine signet ring cell cancers, the low partization gland cancer of 3 examples, 2 routine undifferentiated carcinomas, 2 routine adenosquamous carcinomas, 1 routine squama type cell cancer), 50 is that the colon cancer tissue sample of Different Individual (comprises 16 routine papillary adenocarcinomas, 13 routine tubular adenocarcinomas, 11 routine myxoadenocarcinomas, 4 routine signet ring cell cancers, the low partization gland cancer of 2 examples, 2 routine undifferentiated carcinomas, 1 routine adenosquamous carcinoma, 1 routine squama type cell cancer); 20 is the normal structure contrast sample of Different Individual, comprises the heart, brain, and muscle, lymph node, prostate, adrenal gland, testis, liver, spleen, thyroid gland, the normal structure at pancreas or other positions also can prepare the tissue microarray biochip that other types are arranged.Carry out alkaline phosphatase enzyme staining according to conventional method.Sample must be fixed, wash in 4 ℃ of refrigerators before enzyme dyeing, preserving the activity of enzyme, hatches after 2 hours distillation with 37 ℃ of the effect liquid in the calcium cobalt method and washes 2 times, cleans with distilled water act on 2 minutes in 1% ammonium sulfide after again; Put it into and fully wash after 2% methyl green (through chloroform extraction) is redyed, use the glycerin gelatine sealing.The alkaline phosphatase activities district can be dyed black or brownish black, it is green that nuclear is.If adopt other enzyme stainings also can show various redox enzymes, group's transferase, the activity of hydrolase and synthetic enzyme or content.Be usually used in the diagnosis of enzymatic activity abnormal diseases.Can compare simultaneously with analyze dozens of and even thousands of same type or dissimilar Different Individual or different developing period or other various different tissue samples of arranging in the similarities and differences, variation, and the mutual relationship and the clinical meaning of disease of enzymatic activity.
Embodiment 4: the immunohistochemical analysis of tissue microarray biochip
For the breast cancer tissue's sample P53 albumen of Different Individual and the expression of estrogen receptor (ER) are detected,, and simultaneously a large amount of breast cancer tissue's samples is compared and analyzes so that determine therapeutic scheme and evaluate its prognosis.At first select the tissue specimen of 210 routine Different Individual, comprising the fixing breast cancer tissue of the fresh freezing ethanol of 60 examples, the breast cancer tissue of 120 routine formalin fixed, other normal structure contrasts of the normal breast cancerous tissue of 20 routine Different Individual and 10 routine Different Individual.The histological type of breast cancer comprises 107 routine conduit type, 41 routine microphyll types, and 11 routine marrow templates, 2 routine mucus types, 5 routine tubular-types, 4 routine nipple types were accepted the breast cancer of chemotherapy, 5 routine recurrence breast cancers, 2 routine metastatic carcinomas before the 3 routine arts.PT is the I phase 27% by stages, phase 52%, phase 13%, phase 8%; Nodi lymphatici axillares PN
043%, PN
147%, PN
210%.The normal structure of 10 routine Different Individual comprises brain, the heart, liver, lung, muscle, thyroid gland, prostate, pancreas, stomach and adrenal gland.Equal in a organized way 4 fixedly spend the night, paraffin embedding, section, HE dyeing, back, diagnosis location is prepared into the micro-array tissue piece that each sample diameter is 1mm with mechanization or robotization array preparing instrument, promptly becomes breast cancer tissue's micro-array biochip on the silication slide through cutting into slices and being fixed on.Through dewaxing, dehydration, transparent, carried out antigen retrieval 15 minutes with 90 ℃ of microwaves after the aquation, add the first antibody (DAKO) of anti-p53 albumen or anti-ER, wash after 60 minutes, add the second antibody (antiantibody) of peroxidase labelling again.Add diaminobenzidine colour developing after 60 minutes, with the tumour cell of known stained positive as positive control, with the staining section that do not add first antibody as negative control.If it is promptly positive that the neoplastic cell nuclei more than 10% is obviously dyeed.The SABC method is mainly used in the showed cell nuclear, on the nuclear membrane, and in the tenuigenin, on the cell membrane, in the organelle, nearly all albumen and polypeptide class antigen molecule on the organelle coating and in the tissue.The mark that mainly comprises epithelial tissue and related neoplasms thereof, the label of soft tissue and related neoplasms thereof, the label of lymphohematopoietic tissue delivered and related neoplasms thereof, the antigenic mark of neuroendocrine cell and acceptor thereof, various cell factors and associated receptor thereof, various enzymes and human tissue compatibility antigen and various pathogenic microorganism comprise the specific antigen of bacterium, virus, mould, protozoon, parasite etc.Be usually used in the diagnosis of tumour and non-neoplastic disease, the selection and the prognosis evaluation of antidiastole therapeutic modality, judge the origin and the differentiation of cell, determine the tissue and the interior location of cell of various related antigen molecules, study the correlationship of cellular morphology and function, study the correlationship of various protein moleculars and cell growth, growth, metabolism, differentiation; Whether the content of studying various antigen molecules increases, minimizing or whether the site changes and with the correlationship of cell function and disease.What is more important can compare simultaneously with the tissue samples of analyzing dozens of and even thousands of same type or dissimilar Different Individual or the Different Individual of the Different Individual of same developing period or different developing periods in the mutual relationship and the clinical value of the similarities and differences, variation and disease of same specific molecular or different molecular.
Embodiment 5: the fluorescence in situ hybridization of tissue microarray biochip (FISH) is analyzed
For the amplification situation of specific gene in rapid screening and the different tumor tissues of research, at first prepare the multisample tumor tissues micro-array biochip of Different Individual with mechanization or robotization array preparing instrument.Adopt three kinds of amplification situations that oncogene is Cyclin D1, myc, ErbB-2 in three kinds of different tumor specimens of fluorescein-labeled FISH method analysis.
Micro-array tissue is made up of 500 tissue samples, comprise 15 types 430 routine primary tumo(u)rs and 70 routine normal structure samples, sample is fixed 16 hours for 4 ℃ with freezing ethanol, paraffin embedding, section is dyeed with HE and is diagnosed and take a sample mark, be prepared into the micro-array tissue piece that the sample diameter is 0.6mm with mechanization or robotization array preparing instrument, adopting the paraffin section backup system that 5 μ m array slice thick, 0.6mm is transferred to promptly becomes the tumor tissues micro-array biochip on the silication slide.
Primary tumo(u)r comprises 86 routine breast cancer (31 routine conduit type, 27 routine tubular-types, 23 routine glue templates), 81 routine lung cancer (31 routine squama cancers, 29 routine gland cancer, 9 routine large cell carcinomas, 6 routine small cell carcinomas, 3 routine bronchovesicular cancers), 73 routine sdenocarcinomas of stomach, 39 routine oophoromas (19 routine slurry types, 17 routine mucus types, 3 examples are model in utero), 23 example endometrial carcinomas, 23 routine colorectal cancers, 17 routine incidence cancer (15 routine laryngocarcinoma, 2 routine oral squamous cell carcinomas), 15 routine clear cell carcinomas, 14 routine orchioncus (10 routine spermatogonium cancers, 4 routine teratomas), 16 routine TCCBs, 13 routine thyroid cancers, 15 routine prostate cancers, (3 examples derive from lung to 6 routine carcinoids, 3 examples derive from small intestine), 5 routine chromaffin cell cancers, 4 routine malignant mela nomas.Derive from mammary gland, lung, prostate, pancreas, small intestine, stomach, colon, kidney, liver, thyroid gland, the normal structure of bladder is in contrast.
Before the hybridization, tissue microarray biochip go ahead of the rest dewaxing, dehydration, transparent, aquation are at first carried out pre-service to reduce the background colour developing with Rnase and proteinase, embathed 5 minutes with 37 ℃ of 2 * SSC, added 37 ℃ of 100 μ l RNase 1 hour, with 2 * SS2 rinsing 2 times, PBS rinsing 1 time, 37 ℃ 5 minute/time, the immobile liquid room temperature is fixed 10 minutes, PBS rinsing 2 times, 5 minutes/time, degree of passing ethanol dehydration is dried.Added 80 ℃ of 100 μ l sex change liquid 2-5 minute, 2 * SSC rinsing 5 minutes, degree of passing ethanol dehydration is hybridized after drying.As probe, green glow mark centromeric probe (CEP11 and CEP17) is as reference with Cyelin D1, the myc of orange signal and ErbB-2; 75 ℃ of sex change 5 minutes add hybridization buffer 100 μ l, wherein contain 30ng probe and 15ng people Cot1-DNA, hybridize in 37 ℃ of wet boxes and spend the night, with the 37 ℃ of rinsings of rinsing liquid 3 times that are preheated to 37 ℃, 5 minutes/time, dye with 125 μ g/ml DAPI lining, cover with cover glass, 4 ℃ of magazines are preserved.The FISH signal adopts the BX-51 fluorescent microscope of being furnished with dichroic filter to observe green glow and orange light signal simultaneously.The result is judged to be at least 5% tumour cell FISH signal more than 10; Or wherein 5 signals are the high light signal; Or all be considered as target gene amplification more than 3 times greater than kinetochore signal on the autosome.Found that Cyclin D1 gene magnification is common in mammary gland (26%), lung (10%), neck (41%), and the tumour and the melanoma (25%) at bladder positions such as (13%); ErbB-2 gene magnification is common in bladder (13%), mammary gland (10%), colon (3%), stomach (19%), testis (7%), the tumour at lung positions such as (6%); Myc gene magnification is common in mammary gland (12%), colon (3%), kidney (7%), lung (9%), neck (6%), bladder (16%), ovary (8%) and endometrium (18%), the tumour at testis positions such as (7%).Wherein there are 3 routine breast cancer Cyclin D1 and myc gene to increase simultaneously; 2 routine breast cancer, 1 routine teratoma of testis and 1 routine lung cancer Cyclin D1 and ErbB-2 gene increase simultaneously; 1 routine breast cancer myc and ErbB-2 gene increase simultaneously.Three kinds of oncogenes are increased simultaneously is detected in 1 routine breast cancer.Find that simultaneously these three kinds of oncogenes there is no amplification in normal structure.But because DNA, the RNA of the multiple tissue of tissue microarray biochip technology one-time detection and the multiple target spot of protein molecular, so it is big to have capacity, robotization to advantages such as donor tissue piece damage are little, can accurately be located and is used for different molecules and immunostaining tissue specimen.According to the thickness of original structure piece, can be made into 150-300 and open continuous tissue microarray biochip, can carry out stereoscopic analysis to small tumor specimen like this, kept the tumor specimen of rare preciousness.This technology not only is applicable to various neoplastic disease, also is applicable to every other disease simultaneously, comprises the various diseases of animal; Be applicable to all oncogenes, the intersection of the amplification of tumor suppressor gene or other diseases related gene, variation, inactivation etc. detects, the detection of the various combination of various diseases and normal structure.
Embodiment 6: tissue microarray biochip is used for the detection of tumor tissues PDGF gene
Adopt PDGF (PDGFB) gene in the kinds of tumors micro-array tissue sample in the identical FISH process analysis example 5, PDGFB high level amplification as a result is detected in the malignant tumour of particular type, as breast cancer (3.7%), lung cancer (19.2%), colon cancer (3.2%), carcinoma of testis (7.1%) TCCB (37.5%) and carcinoma of endometrium (28.6%), and other malignant tumour and normal control all do not have the PDGFB amplification.Since this method can be in each micro-array tissue the hundreds of and even thousands of samples of high-throughout analysis, promptly can be used for DNA analysis, also can be used for the analysis and the location of RNA and protein molecular.Because the characteristics of its technology itself, can study same tumor types or other diseases simultaneously, also can study dissimilar tumours or other diseases simultaneously, or while progress phase tumour, the paracmasis tumour, the treatment pre-neoplastic, treatment back tumour, or study simultaneously same tumor types tens to a hundreds of Prognosis molecular marker.And each the cylindric tissue specimen in the micro-array tissue wax stone also can be used as the deposit of DNA and RNA molecule, to its extract, separation and purifying can be used for further molecular biological analysis.
Embodiment 7: tissue microarray biochip is used for studying the amplification of prostate cancer evolution several genes
Mode by embodiment 1 prepares the prostate cancer tissue micro-array biochip, relates to different prostate cancer sample and the 32 routine metastatic carcinoma samples of 112 examples altogether; 31 routine benign prostatic hyperplasis samples (BPH) in contrast, normal structure contrasts with embodiment 1.Double-colored FISH method adopts the plain mark AR of fluorescent orange, c-myc, ErbB-2, four kinds of gene probes of Cyclin D1, and FITC (fluorescein isothiocynate) mark centromeric probe in contrast.Monochromatic FISH adopts the plain mark n-myc of fluorescent orange gene probe.Operation is undertaken by embodiment 5.The gene magnification standard is that fluorescence signal in 10% tumour cell is greater than contrast probe (centromere) more than 3 times; The ratio of to be measured and crt gene fluorescence probe counting can not can only be considered as low-level amplification as the foundation of gene specific amplification between 1-3 the time; The standard of anecdotal n-mcy gene magnification is the fluorescence signal that has in the tumour cell more than 10% more than 5.
Analysis result shows, the amplification of X chromosome AR gene in 96% the BPH sample, 84% No. 8 chromosome c-myc genes, No. 17 chromosome ErbB2 genes of 81% and 83% No. 11 chromosome cyclin D1 genes is all with centric hyperfluorescence signal.To have the average percent of two kinds of fluorescence signals be 75% to autosome in epithelial cell, and having only a kind of fluorescence signal is 20%, no fluorescence signal only account for 5%.In addition, 24.0% has AR gene magnification in the invalid recurrent lesions of 33 routine hormone therapies, and 25% has AR gene magnification in the invalid metastatic carcinoma of 32 routine hormone therapies.In 112 routine primary tumo(u)rs, only there is 1 example (1%) that AR gene magnification is arranged, and no AR gene magnification in the 31 routine BPH contrasts.Show that prostate cancer and AR gene magnification that hormone therapy is invalid are closely related.And, if AR gene magnification is arranged, the amplification of AR gene must be arranged also in the then corresponding metastatic carcinoma in the primary tumo(u)r.In metastasis 11% and 5% the recurrent focus, high-level c-myc gene magnification is arranged all; But in 112 routine primary lesions and 31 routine BPH contrasts, all there is not amplification.In the tumor specimen of 22 routine AR amplification, 12% c-myc gene magnification is arranged simultaneously in addition, and in the sample of the no AR gene magnifications of 96 examples only 2% sample c-myc gene magnification is arranged.Prove absolutely that AR and c-myc gene have tangible correlativity.
In primary tumo(u)r, only 2% has cyclin D1 gene magnification, and in recurrent tumor up to 9%, be 5% in metastasis.In the tumour of 60% cyclin D1 amplification without the amplification of other genes.The amplification of ErbB-2 gene and n-myc gene is seldom arranged in the prostate cancer, also do not have ErbB-2 gene magnification among the BPH.
Above-mentioned experimental result shows that the growth of the tumour cell that hormone therapy is invalid and AR gene magnification are closely related, c-myc gene magnification then with shift relevant.The amplification of AR gene is the most common in prostate cancer, often is independent of c-myc and cyclinD1.Accidental cyclin D1 gene magnification, and ErbB-2 and n-myc gene all do not have amplification any period in prostate cancer.
Embodiment 8: the tissue microarray biochip technology is in conjunction with cDNA microarray technology rapid screening clear-cell carcinoma (RCC) prognostic marker: at first use the gene that plays an important role among the cDNA microarray technology identification RCC, further analyze the possible clinical meaning of these genes by tumor tissues micro-array biochip technology then.
Adopt the standardization experimental technique, extract total RNA of normal kidney and renal carcinoma cell line CRL-1933, synthetic cDNA also uses fluorescence labeling, hybridizes and analyze the mode of 5200 kinds of gene expressions of normal kidney tissue and CRL-1933 with the cDNA microarray of the known of 5200 dot matrix or expressed sequence (EST).Low or the high expressed of certain gene or EST need satisfy following two conditions: 1. normal kidney tissue and CRL-1933 express and differ more than 10 times, and 2. visual inspection cDNA dot matrix filter membrane has marked difference.It is different that analysis result finds to have 89 genes or EST to express.There are 38 sequences to be high expressed among the CRL-1933, comprise 26 known genes and several EST; 51 sequences are low to be expressed, and comprises 25 known genes and 26 est sequences.In tumour cell, the gene order of one of them rise is identical with the sequence of vimentin.
The structure of kidney neoplasms tissue microarray biochip is with embodiment 1.Comprise 250 kinds of tumor of kidney samples and 30 routine normal kidney tissue samples.The IIP method of employing standard is carried out immunohistochemical staining.One anti-is the vimentin monoclonal antibody, with the positive tissue of endothelial cell vimentin as positive control, not add a tissue specimen that resists as negative control.Have tangible cell dyeing to be the vimentin positive in the tumour cell, the dye-free person is then negative.Found that 232 examples (91%) tumor tissues sample and 30 routine normal kidney tissue sample vimentins are positive, its expression is more common in and is non-ly had a liking among the look RCC, expresses seldom and have a liking among the look RCC.Normal renal tubule is not expressed vimentin.In non-dyeing RCC, the expression of its vimentin is increased with TNM by stages.Through having a liking for look patient RCC and follow up a case by regular visits to (average 52 months) and find that vimentin is expressed strong more non-, patient is short more life cycle.Show that the vimentin expression can be used as an independently prognosis judgement symbol.In addition, to have a liking among the look RCC positive rate higher non-for vimentin, and comparatively rare in having a liking for look RCC, so vimentin also can be used as the index of both antidiastoles.
The above results shows, tumor tissues micro-array biochip technical tie-up cDNA microarray technology all plays crucial effect in the judgement of gene expression, genescreen, the assignment of genes gene mapping, gene function and clinical value in identification and assessment human malignancies.If adopt traditional pathology test method, finishing this process approximately needs the time of several years, and tumor tissues micro-array biochip technology be owing to can analyze DNA, RNA and protein in hundreds and thousands of different tumor tissues samples simultaneously, thereby shortened the search time of job family greatly.In addition, the achievement that tumor tissues micro-array biochip technology can be quickened in the fundamental research transforms to clinical practice, because synchronous express-analysis is achieved in the research of different step, as in dissimilar kinds of tumors sample dot matrix, carrying out earlier the screening and the detection of specific gene or molecular marker, as find specific gene or molecular marker high expressed in specific tumors, then further analyze by the specific tumors micro-array tissue again or carry out perspective study by the microarray immunohistochemistry technique, and estimate its clinical value, thereby it is the achievement of fundamental research was shortened dramatically to the time that clinical practice transforms, and easy, economical.As blood platelet derivation growth factor B (PDGFB) among the breast carcinoma cell strain SKBR3 being detected by said method, dna microarray detects finds that the PDGFB amplification is arranged among the SKBR3, utilize this discovery to prepare the PDGFB probe, be used to screen breast cancer tissue's microarray, found that only 2% breast cancer has the PDGFB amplification.Yet find that then the PDGFB gene is high expressed in lung cancer and carcinoma of urinary bladder when adopting this probe in detecting kinds of tumors micro-array tissue, so just found a new diagnosing tumor mark.
Embodiment 9: the tissue microarray biochip technology is used for the analysis that gene expression frequencies, distribution and cancer development process gene change
The tissue microarray biochip technology can be used for the research of a large amount of genes, and in conjunction with the cDNA array technique, the pattern of contrast gene expression can be understood tumorigenic molecule mechanism better and find that new examining in advance indicates and potential treatment target spot.
In order to study insulin-like growth factor binding protein 2 (IGFBP2) and the relation that the human prostate cancer makes progress, at first adopt the gene IGFBP2 of cDNA microarray technology screening unconventionality expression, make up the prostate cancer tissue micro-array biochip then.Select formalin fixed and paraffin-embedded 96 routine primary prostate cancers, the tissue specimen of 39 routine local recurrence prostate cancers, with the tissue specimen of 31 routine benign prostatauxes in contrast.The IIP method of employing standard is carried out immunohistochemical staining.Repair antigen with microwave, one anti-(C-18) is sheep polyclonal antibody IGFBP2, and reaction develops the color with DAB.The IGFBP2 positive control is a kidney cortex, does not add a sample that resists as negative control.The cytoplasm tinter is positive.Found that the positive rate in the non-hormone tolerance primary tumor is 30%, the positive rate in the recurrent tumor is up to 96%, and the normal structure contrast is negative.Show that the suspicious gene of being found through the cDNA microarray hybridization can directly confirm through the tissue microarray biochip technology.The above results shows that IGFBP2 can be used as progressive stage prostate cancer diagnosis and the index of judging prognosis, also can be used as the new target spot of treatment tumour.
Embodiment 10: the tissue microarray biochip technology can be used for determining the tumor treatment scheme
The tissue microarray biochip technology can be used for screening gene expression relevant with treatment in a spot of tumor tissues sample.As breast cancer tissue's sample is carried out the HER-2 genescreen, the patient with breast cancer of high expressed can treat with specific antibody (Herceptin).Herceptin can with the HER specific bond, thereby suppress the HER-2 expression of gene, so have excellent curative.This information can be used for also judging whether other tumours can be used for the treatment of Herceptin.
Embodiment 11: the tissue microarray biochip technology is used for the screening of tumor prognosis mark
Make up known medical history and result's patient's tumor tissue microarray biochip, it is very useful being used to screen prognostic marker.Adopt original position RT-PCR technology, immunohistochemistry technique and restriction fragment length polymorphism analytical technology to detect tissue specimen in the various known tumor tissues microarray, analysis result shows that the oophoroma of 40% breast cancer, 60% colon cancer, 60% carcinoma of urinary bladder, 40% cancer of the esophagus, 37% cancer of the stomach and 50% all has sudden change, inactivation or the disappearance of P53 gene, and the high mutation rate of P53 and patient's poor prognosis are closely related.
Embodiment 12: the tissue microarray biochip technology is used to seek the target spot of new gene therapy
It is highly effective that the tissue microarray biochip technology is used to seek the new target gene for the treatment of tumour.On the tissue microarray biochip that contains thousands of different tumor samples, adopt different probes to detect, to find new proto-oncogene or the gene of the new signal transducers of encoding.Such probe can react with one or more tumours.If probe discloses certain special gene overexpression or express enhancing in many tumours, then this gene just becomes a kind of crucial target gene.This target gene plays an important role in different tumours of the same race or different types of tumors, and the material that suppresses this expression of gene so or suppress the function of the expressed product of this gene may be exactly the new drug that has potentiality.Show that it is crucial that tumor tissues micro-array biochip technology is at first screened target gene when developing new cancer therapy drug.
Embodiment 13: the application of tissue microarray biochip technology in clone
The cell of in vitro culture or from non-solid tissue such as blood, marrow, fine needle aspiration sample, hydrothorax, ascites isolated cells also can be used for making tissue microarray biochip analysis, this is that the tissue microarray biochip technology is used for the impressive progress that cell individual is analyzed, and this will make the flow of research of non-solid tumours such as leukaemia, lymthoma and myeloma greatly accelerate.
With in vitro culture or from other body fluid isolated cells or clone, with making cell suspension after the trypsin treatment, 1200G is centrifugal, the gained cell precipitation is fixed with formaldehyde-ethanol matrix, the routine paraffin wax embedding comprises the tens of kinds of cell microarray biochips to thousands of kinds of different cell colonys by the preparation of the method for above-mentioned making tissue microarray biochip.This cell microarray biochip can be analyzed with different modes, not only can also can analyze function, distribution, the expression of special gene with unusual cell specimen with normal.
Normal tissue specimen comprises health adult tissue, embryo's sample tissue, the health adult tissue of different developmental phases, intact animal tissue, the animal tissue of different developmental phases, animal embryo tissue and various transgenic animals tissue.
The micro-array tissue technology not only can be used for analyzing neoplastic disease and genetic disease, also is applicable to the non-genomic disease.If find that in analytic process certain gene or protein expression mode are unusual to some extent, this unusual generation and diagnosis to disease may be vital.
Yeast or bacterium also can be made into cell suspension, can be made into solid or semisolid fritter behind the cell centrifugation in the suspension, fixing then and preparation cell microarray biochip.The liquid cell suspension is made the cell microarray biochip among also can importing matrix by conduit, as be incubated at the cell suspension that chemotherapy detects that is used in the microtest plate, cell in each hole is accepted the medicine of different types of medicine or variable concentrations, successively observes and analyze the expression of function, feature, form, survival rate and the special gene of each clone in the implantation cell microarray under the medicine effect.The application cell microarray technology can carry out histology or immunology research, also can unrestrictedly be used for nucleic acid hybridization, original position PCR, PRINS, ligase chain reaction or the like.
The tissue microarray biochip technology also can be used for the detection of biopsy specimen except that above application, as operation biopsy, aspiration biopsy, various endoscopic biopsy sample.These samples can directly be fixed, and are prepared into the biopsy specimen tissue microarray biochip, use for clinical or research.As use various molecular marked compound to predict the prognosis and the development of various precancerous lesions, perhaps in transfer that can not excision or recurrent tumor, detect to the reaction of treatment with to the prediction of result of treatment and patient's prognosis evaluation etc.
In addition, cell in the haemocyte of cervical smear, blood preparation, separation, arena cell, the ascites in cell, the various transudate, sputum cells etc. also can be made various cell microarray biochip after centrifugal, fixing, this not only provides a kind of cytodiagnosis method fast, and can carry out manual, semi-automatic or automatic analysis to various cellular morphology and characterization of molecules simultaneously.This method is very important for the analysis and the diagnosis of leukaemia, lymthoma, pernicious ascites pleural fluid and bloody sputum etc.Can carry out simultaneously immunophenotype, the viral antigen of many samples, the detection and the analysis of viral DNA or other pathology factors.
The tumor tissues micro-array biochip of different stages of development can reflect the development process of tumour, can diagnose antidiastole, treatment monitoring and prognosis evaluation to tumour by analyzing specific biochemical molecular or gene expression.
Obtain the biological sample of sudden change or particular model at the different times that grows from animal pattern or transgenic animals; or obtain biological sample from the budding embryonic tissue of difference and postnatal different developmental phases and carry out various analyses, thereby the molecular difference of gene expression in the more normal or unusual developing embryo process.
Utilize the tissue microarray biochip technology to carry out all analyses to all cells of all animals (comprising the people).
Claims (8)
1. tissue microarray biochip, comprise and derive from various malignant tumours, various benign tumours, various precancerous lesions, various non-neoplastic diseases, various normal structures, various embryonic tissues, various blood and hematopoietic tissue, various cell extracts, various biopsies or cell, various hydrothorax, ascites, urine, sputum, cell in the transudate, the histocyte of various in vitro culture and clone, various bacteriums and saccharomycete, the normal structure of various animals, the pathological tissues of various animals, the embryonic tissue of various animals, various animal model tissues, the donor paraffin organization sample of various transgenic animals tissues or various transgenosis organs, it is characterized in that: adopt mechanization apparatus for preparation or automated preparation instrument from donor paraffin organization sample, to take out cylindric tissue core, each cylindric tissue core is placed in the blank wax stone of another piece (acceptor piece) by design sequence, formative tissue microarray wax stone, from this micro-array tissue wax stone, can obtain a large amount of micro-array tissue sections, section being placed on the slide of silication and gel is the formative tissue micro-array biochip, comprises a large amount of orderly various donor paraffin organization samples on each tissue microarray biochip.
2. tissue microarray biochip according to claim 1 is characterized in that: described donor paraffin organization sample all will be through the privileged site of drawing materials, observation under ethanol or formaldehyde fixed, dehydration, transparent, waxdip, embedding, section, HE dyeing or other specific stains, the optical microscope, diagnosis and position tissue sample will be studied and take a sample.
3. tissue microarray biochip according to claim 1 is characterized in that: described cylindric tissue core of being got from the donor paraffin organization sample, and its diameter is 0.2mm～4.0mm, that commonly used is 0.5mm～2.0mm; The height of cylindric tissue core is 1.0mm～8.0mm, and often the height that adopts is 2.0mm～5.0mm.
4. tissue microarray biochip according to claim 1 is characterized in that the area of described every kind of micro-array tissue wax stone is respectively 15mm * 15mm, 15mm * 20mm, 15mm * 25mm, 15mm * 30mm, 15mm * 35mm, 15mm * 40mm, 15mm * 45mm, 15mm * 50mm, 20cm * 15mm, 20mm * 20cm, 20cm * 25cm, 20cm * 30cm, 20cm * 35cm, 20cm * 40cm, 20cm * 45cm, 20cm * 50cm; 25cm * 15mm, 25mm * 20mm, 25mm25cm, 25cm * 30cm, 25cm * 35cm, 25cm * 40cm, 25cm * 45cm, 25cm * 50cm.
5. according to claim 1 or 4 described tissue microarray biochips, it is characterized in that the thickness of described every kind of micro-array tissue wax stone is respectively 2.0mm, 3.0mm, 4.0mm, 5.0mm, 6.0mm, 7.0mm, 8.0mm, 9.0mm, 10.0mm, 11.0mm, 12.0mm, 13.0mm, 14.0mm, 15.0mm, 16.0mm, 17.0mm, 18.0mm, 19.0mm, 20.0mm.
6. tissue microarray biochip according to claim 1, it is characterized in that to place on every tissue microarray biochip a tissue array of different sizes, or place big or small on an equal basis or two big or small tissue arrays of difference, or place three tissue arrays of equal size or different sizes, or place four tissue arrays of equal size or different sizes.
7. according to claim 1 or 3 described tissue microarray biochips, it is characterized in that: describedly can from donor paraffin organization sample, take out cylindric tissue core, diameter is the cylindric tissue core of 0.2mm, the micro-array tissue that can prepare 2～8000 samples on a slide, preferable array number are 20～5000 tissue samples; Taking out diameter is the cylindric tissue core of 0.3mm, can prepare the micro-array tissue of 2～5000 samples on a slide, and preferable array number is 20～2180 tissue samples; Taking out diameter is the cylindric tissue core of 0.4mm, can prepare the micro-array tissue of 2～3000 samples on a slide, and preferable array number is 20～1250 tissue samples; Taking out diameter is the cylindric tissue core of 0.5mm, can prepare the micro-array tissue of 2～2000 samples on a slide, and preferable array number is 20～800 tissue samples; Taking out diameter is the cylindric tissue core of 0.6mm, can prepare the micro-array tissue of 2～1500 samples on a slide, and preferable array number is 20～640 tissue samples; Taking out diameter is the cylindric tissue core of 0.7mm, can prepare the micro-array tissue of 2～1200 samples on a slide, and preferable array number is 20～520 tissue samples; Taking out diameter is the cylindric tissue core of 0.8mm, can prepare the micro-array tissue of 2～960 samples on a slide, and preferable array number is 20～450 tissue samples; Taking out diameter is the cylindric tissue core of 0.9mm, can prepare the micro-array tissue of 2～800 samples on a slide, and preferable array number is 20～390 tissue samples; Taking out diameter is the cylindric tissue core of 1.0mm, can prepare the micro-array tissue of 2～640 samples on a slide, and preferable array number is 20～330 tissue samples; Taking out diameter is the cylindric tissue core of 1.1mm, can prepare the micro-array tissue of 2～520 samples on a slide, and preferable array number is 20～300 tissue samples; Taking out diameter is the cylindric tissue core of 1.2mm, can prepare the micro-array tissue of 2～450 samples on a slide, and preferable array number is 20～270 tissue samples; Taking out diameter is the cylindric tissue core of 1.3mm, can prepare the micro-array tissue of 2～390 samples on a slide, and preferable array number is 20～240 tissue samples; Taking out diameter is the cylindric tissue core of 1.4mm, can prepare the micro-array tissue of 2～330 samples on a slide, and preferable array number is 20～210 tissue samples; Taking out diameter is the cylindric tissue core of 1.5mm, can prepare the micro-array tissue of 2～300 samples on a slide, and preferable array number is 20～200 tissue samples; Taking out diameter is the cylindric tissue core of 1.6mm, can prepare the micro-array tissue of 2～270 samples on a slide, and preferable array number is 20～180 tissue samples; Taking out diameter is the cylindric tissue core of 1.7mm, can prepare the micro-array tissue of 2～240 samples on a slide, and preferable array number is 20～160 tissue samples; Taking out diameter is the cylindric tissue core of 1.8mm, can prepare the micro-array tissue of 2～210 samples on a slide, and preferable array number is 20～140 tissue samples; Taking out diameter is the cylindric tissue core of 1.9mm, can prepare the micro-array tissue of 2～200 samples on a slide, and preferable array number is 20～130 tissue samples; Taking out diameter is the cylindric tissue core of 2.0mm, can prepare the micro-array tissue of 2～170 samples on a slide, and preferable array number is 20～125 tissue samples; Taking out diameter is the cylindric tissue core of 2.1mm, can prepare the micro-array tissue of 2～160 samples on a slide, and preferable array number is 20～120 tissue samples; Taking out diameter is the cylindric tissue core of 2.2mm, can prepare the micro-array tissue of 2～140 samples on a slide, and preferable array number is 20～110 tissue samples; Taking out diameter is the cylindric tissue core of 2.3mm, can prepare the micro-array tissue of 2～130 samples on a slide, and preferable array number is 20～105 tissue samples; Taking out diameter is the cylindric tissue core of 2.4mm, can prepare the micro-array tissue of 2～125 samples on a slide, and preferable array number is 20～95 tissue samples; Taking out diameter is the cylindric tissue core of 2.5mm, can prepare the micro-array tissue of 2～120 samples on a slide, and preferable array number is 20～90 tissue samples; Taking out diameter is the cylindric tissue core of 2.6mm, can prepare the micro-array tissue of 2～110 samples on a slide, and preferable array number is 20～85 tissue samples; Taking out diameter is the cylindric tissue core of 2.7mm, can prepare the micro-array tissue of 2～105 samples on a slide, and preferable array number is 20～80 tissue samples; Taking out diameter is the cylindric tissue core of 2.8mm, can prepare the micro-array tissue of 2～95 samples on a slide, and preferable array number is 20～75 tissue samples; Taking out diameter is the cylindric tissue core of 2.9mm, can prepare the micro-array tissue of 2～90 samples on a slide, and preferable array number is 20～70 tissue samples; Taking out diameter is the cylindric tissue core of 3.0mm, can prepare the micro-array tissue of 2～85 samples on a slide, and preferable array number is 20～65 tissue samples; Taking out diameter is the cylindric tissue core of 3.1mm, can prepare the micro-array tissue of 2～80 samples on a slide, and preferable array number is 20～60 tissue samples; Taking out diameter is the cylindric tissue core of 3.2mm, can prepare the micro-array tissue of 2～75 samples on a slide, and preferable array number is 20～55 tissue samples; Taking out diameter is the cylindric tissue core of 3.3mm, can prepare the micro-array tissue of 2～68 samples on a slide, and preferable array number is 20～52 tissue samples; Taking out diameter is the cylindric tissue core of 3.4mm, can prepare the micro-array tissue of 2～65 samples on a slide, and preferable array number is 20～48 tissue samples; Taking out diameter is the cylindric tissue core of 3.5mm, can prepare the micro-array tissue of 2～60 samples on a slide, and preferable array number is 20～44 tissue samples; Taking out diameter is the cylindric tissue core of 3.6mm, can prepare the micro-array tissue of 2～56 samples on a slide, and preferable array number is 20～40 tissue samples; Taking out diameter is the cylindric tissue core of 3.7mm, can prepare the micro-array tissue of 2～52 samples on a slide, and preferable array number is 20～36 tissue samples; Taking out diameter is the cylindric tissue core of 3.8mm, can prepare the micro-array tissue of 2～48 samples on a slide, and preferable array number is 20～32 tissue samples; Taking out diameter is the cylindric tissue core of 3.9mm, can prepare the micro-array tissue of 2～44 samples on a slide, and preferable array number is 20～28 tissue samples; Taking out diameter is the cylindric tissue core of 4.0mm, can prepare the micro-array tissue of 2～40 samples on a slide, and preferable array number is 20～24 tissue samples.
As claimed in claim 1, wherein the tissue microarray biochip , characterized in that the tissue microarray chip set
Woven lattice order is pre-set , arranged according to the following combinations : 1 ) . Arranged in the same disease : about the same disease
Tissue samples from different individuals as well as a variety of different individuals are arranged in the normal control tissue samples a tissue microarray biochips
On ; 2 ) . Different diseases arranged : Upcoming different diseases as well as tissue samples from different individuals in a variety of different individuals normal controls
Specimens arranged in a tissue microarray chip ; 3 ) The same disease the same site Arrangement: about the same organ lesions
Weaving , the lesion tissue , normal tissue as well as a variety of different individuals are arranged in the normal control tissue samples a tissue microarray
Chip ; 4 ) . Ordered the same parts of different diseases : Upcoming different organs of diseased tissue , the lesion tissue , normal tissue and non-
Individuals with a variety of normal control tissue samples arranged in a tissue microarray chip ; 5 ) The same disease the same tissue type
Ordering : Coming same disease the same type of tissue specimens of different individuals and different individuals of various normal control tissue samples
The arranged in a tissue microarray biochips ; 6 ) . Different diseases the same organization Genre : Coming same group of different diseases
Different types of organizations as well as individual specimens of a variety of different individuals are arranged in the normal control tissue samples a tissue microarray
Chip ; 7 ) The same disease the same clinical stage arrangement : Coming same disease with a clinical staging tissue samples of different individuals
As well as a variety of different individuals are arranged in the normal control tissue samples a tissue microarray chip ; 8 ) . Different diseases the same Pro
Bed installment arrangement : Coming different clinical stages of the disease the same kind of tissue samples from different individuals as well as a variety of different individuals in control
Tissue samples arranged in a tissue microarray chip ; 9 ) . Ordered the same sex the same disease : the same about the same disease
Kinds of tissue samples from individuals of different genders and a variety of different individuals are arranged in the normal control tissue samples a tissue microarray
Chip ; 10 ) . Different diseases are arranged the same sex : sex is about the same kind of different diseases, tissue samples of different individuals and different
A variety of individual normal control tissue samples arranged in a tissue microarray chip ; 11 ) The same disease the same age row
Column : Coming same disease the same age as well as tissue samples from different individuals in a variety of different individuals normal control tissue samples
Arranged in a tissue microarray chip ; 12 ) . Different diseases are arranged the same age : Age is about the same kind of different diseases
Segments of different tissue samples from individuals as well as a variety of different individuals are arranged in the normal control tissue samples a tissue microarray biochips
On ; 13 ) . Treat the same disease in tandem : Coming same disease before treatment or after treatment tissue samples of different individuals and non-
Individuals with a variety of normal control tissue samples arranged in a tissue microarray chip ; 14 ) . Before or after the treatment of different diseases
Ordering : Coming treatment of different diseases before or after treatment, tissue samples of different individuals as well as a variety of different individuals normal control tissue samples
The arranged in a tissue microarray biochips ; 15 ) . Arranged in the same disease the same treatment : about the same disease the same
Kind of treatment as well as tissue samples of different individuals a variety of different individuals are arranged in the normal control tissue samples a tissue microarray
Biochip ; 16 ) . Different diseases are arranged the same treatment : about the same kind of treatment of different diseases in different ways individual organizations
Specimens as well as a variety of different individuals are arranged in the normal control tissue samples a tissue microarray chip ; 17 ) The same disease
The same arrangement of molecular markers : Coming same disease contain the same molecular markers of tissue samples of different individuals and different individuals and more
Kind of normal control tissue samples arranged in a tissue microarray chip ; 18 ) of different molecular markers arranged in the same disease : the
The molecular markers of different diseases with different individuals of the same tissue samples as well as a variety of different individuals are arranged normal control tissue samples
In a tissue microarray chip ; 19 ) The same disease primary, secondary ( metastasis ) , recurrent lesions arranged : Coming same
Diseases of different individuals or recurrent primary or secondary tumor tissue samples and a variety of different individuals normal control tissue specimens row
Listed in a tissue microarray chip ; 20 ) . Different diseases, primary, secondary ( metastasis ) , recurrent lesions arranged : coming not
Different individuals with the disease, primary or secondary or recurrent tumor tissue samples as well as a variety of different individuals of normal control tissue specimens row
Listed in a tissue microarray chip ; 21 ) . Origins same organization arranged the same disease : about the same group of the same disease
Woven tissue of origin as well as different individuals a variety of different individuals are arranged in the normal control tissue samples a tissue microarray
Chip ; 22 ) . Different diseases are arranged the same tissue origin : Coming same tissue origin of different diseases tissue samples of different individuals
As well as a variety of different individuals are arranged in the normal control tissue samples a tissue microarray chip ; 23 ) . The same disease the same
Differentiation arrangement : the same kind of disease is about the same degree of differentiation of tissue samples from different individuals and different individuals of various normal
Control tissue specimens arranged in a tissue microarray chip ; 24 ) . Different diseases are arranged the same degree of differentiation : Upcoming different
Diseases same degree of differentiation of individual tissue samples and a variety of different individuals normal control tissue samples arranged in a group
Weave microarray chip ; 25 ) . Hierarchical arrangement of the same disease the same : about the same disease classification of different individuals of the same organization
Specimens as well as a variety of different individuals are arranged in the normal control tissue samples a tissue microarray chip ; 26 ) . Different diseases
Same hierarchical arrangement : the same about different diseases classification and tissue samples of different individuals a variety of different individuals normal controls
Specimens arranged in a tissue microarray chip ; 27 ) . Ordered the same system : about different diseases within the human body is not the same system
Individual tissue samples with different individuals as well as a variety of normal control tissue samples arranged in a tissue microarray chip ;
28 ) Multi- system arrangement : Coming human multi-system disease in different tissues of different individuals and different individuals on a variety of normal
According to the tissue specimens arranged in a tissue microarray chip ; 29 ) . History of the same disease the same arrangement : Coming same kind of illness
Disease, a history of different individuals with the same tissue samples and a variety of different individuals normal control tissue samples arranged in a tissue
Microarray chip ; 30 ) . History of the same disease different arrangement : about the same disease , a history of different individuals with different
Tissue samples as well as a variety of different individuals are arranged in the normal control tissue samples a tissue microarray chip ; 31 ) . With
An infectious disease, the same type of arrangement : Coming with a disease that has the same type of infection and tissue samples of different individuals
A variety of different individuals are arranged in the normal control tissue samples a tissue microarray chip ; 32 ) . Different from the same disease infection
Type of arrangement : Coming infected with a disease of different types of tissue samples from different individuals and different individuals on a variety of normal
According to the tissue specimens arranged in a tissue microarray chip ; 33 ) . Same organ normal tissue array : Coming body the same device
Official tissue samples of different individuals are arranged in a tissue microarray chip ; 34 ) . Different organs normal tissue array : Coming
Different organs of the body tissues of different individuals are arranged in a tissue microarray chip ; 35 ) . Same organ the same developmental
Stages of embryonic tissue arrangement : the same about the same developmental stage embryonic organ tissue samples from different individuals and different individuals adults
A variety of normal control tissue samples arranged in a tissue microarray chip ; 36 ) . Different organs of the same developmental stage embryo group
Woven arrangement : Coming same developmental stage embryonic tissues of different individuals in different organs of adult tissue samples and a variety of different individuals are
Normal control tissue samples arranged in a tissue microarray chip ; 37 ) The same organs in different developmental stages of embryonic tissues row
Column : Coming same organ at different developmental stages of embryonic tissue as well as tissue samples of different individuals in a variety of different individuals normal for adults
According to the tissue specimens arranged in a tissue microarray chip ; 38 ) . Different organs at different developmental stages of embryonic tissue arrangement : the
Different organs in different developmental stages of embryonic tissue as well as tissue samples from different individuals in a variety of different individuals were normal adult tissues
Specimens arranged in a tissue microarray chip ; 39 ) Animal tissue arranged : including a variety of different animals in different disease
Disease, normal tissues of different organs in different developmental stages of embryonic tissue , a variety of transgenic animal organs and tissues in different months
The body tissue , the basic arrangement of the above ( 1 to 34 ) arranged in the same human tissue specimens ; 40 ) . Same cell lines with the tissue
Ordering : Coming with the same kind of tissue specimens in vitro cell lines arranged in a tissue microarray chip ; 41 ) . Isomorphism
Tissue cells arranged : Coming isomorphism tissue specimens cultured cell lines arranged in a tissue microarray chip ;
42 ) . Hybrid aligned with the cell lines : Coming with a variety of different species in vitro cell lines arranged in a tissue microarray raw
Was the chip ; 43 ) . Xenogeneic hybrid cell lines arranged : about different species of different cell lines in vitro are arranged on a
Tissue microarray biochip ; 44 ) . Same type of microorganism Ordering : about the same type of microbial cells arranged in a group
Weave microarray chip ; 45 ) . Different types of microorganisms Arrangement: about the different types of microbial cells arranged in an organization
Microarray chip.
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| CN114018680A (en) * | 2021-10-28 | 2022-02-08 | 上海交通大学医学院附属第九人民医院 | Manufacturing method of choroidal melanoma specimen wax block and chip |
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