CN103347574A

CN103347574A - Cell array quality control device for pathological analysis

Info

Publication number: CN103347574A
Application number: CN2010800349065A
Authority: CN
Inventors: 朱伟星
Original assignee: Individual
Current assignee: Individual
Priority date: 2010-12-07
Filing date: 2010-12-07
Publication date: 2013-10-09
Anticipated expiration: 2030-12-07
Also published as: WO2012078138A1; CN103347574B

Abstract

A cell array quality control device for pathological analysis is disclosed together with methods for making and using the device. Cultured celis of plurality of cancer types and origins are placed on solid substrates in a low density array format constituting a wide panel of positive and negative indicators for immunological and molecular assay procedures such as immunahistochemistry, immunocytochemistry, and in situ hybridization.

Description

The cellular array quality control apparatus of pathology morphological analysis

Technical field

The invention relates to the method for making and using the quality control slide that is loaded with the low-density cellular array.Because the characteristic of the cultured cell on the slide is known maybe can knowing by research, these cells can be used as the positive or negative contrast of histopathology.These pathology detect and comprise immunohistochemistry (IHC), immunocytochemistry (ICC), in situ hybridization (ISH), and CYTOGENETIC ANALYSIS OF ONE.The amount control slide that the tissue that cellular array quality control slide can replace usually being difficult to obtain is made.

Background technology

Pathology detect, and such as IHC, ICC, and ISH are used in and do routine cancer diagnosis and prediction on the patient specimen.General IHC, ICC and ISH detect need advance many steps, comprised the acquisition biological sample, and fixed preparation in formalin is handled sample and is dewatered and transparent purpose to reach, specimen embedding in paraffin, the series section also is stated from the microscope slide.Next also to dewax (at dimethylbenzene, handling in alcohol and the water), finally carry out various reactions.Reaction comprises a series of solution, as enzyme, and first antibody, SA detects reagent, and developer drips on histotomy insulation and wash-out.After reaction was finished, histotomy was placed in microscopically and observes.

What the microexamination of traditional biopsy sample showed is overall cell and organizational structure.For example, at bush commonly used and Yihong dyeing (H﹠amp; E) under, nucleus is by purple (brazilwood dyeing), and cytoplasm is dyed redness (Yihong dyeing).Yet, from H﹠amp; The information that E dyeing draws usually is not enough to do diagnosis accurately.The result that IHC and ISH detect can expand to molecular level to cell size and the shape analysis to tissue samples.Can detect specific albumen and expression of gene in the tissue samples with antibody and nucleic acid probe.Whether some existence that detects target or biomarker can show the pedigree of tumour, helps to diagnose or to the prognosis of specific antitumor therapy.In addition, these methods also can be used for detecting microorganism, as bacterium, and fungi, and virus.

Most of Pathology Labs are made the positive control of IHC with the tissue specimen of determining to contain specific antigen in advance.This need register one group of tissue specimen as the IHC positive control, section and file.When positive tissue samples uses up, need prepare new.Each antibody needs a positive tissue contrast in daily IHC testing process.So each antibody is all verified in a certain concentration.

As a kind of improvement, Battifora (United States Patent (USP) 4,820,504and 5,610,022) has set forth a plurality of positive tissue samples (mostly being the tumor tissues sample) has been embedded in the paraffin mass together, simultaneously method in contrast.This " many tissue tumors wax stone " simplified the slicing processes of positive control tissue.People such as Furmanski have set forth a kind of preparation of diverse ways slightly " many tissue tumors wax stone " (U.S.Pat.No.4,914,022).Their improvement is the core of organizing that cuts out from a plurality of basic stitches with the common beverages suction pipe is embedded in the paraffin mass.

Yet using-system causes very big burden in contrast the histology technician, because must collect unnecessary tissue samples, preparation contrasts slide, and their antigenic characteristic is analyzed.Many researchers used non-organization material, such as cell even the polypeptide of in vitro culture.The manufacturer of many researchers and IHC diagnostic kit contrasts the cell point of known dyeing property on slide as positive or negative.People such as Moskaluk (C.A.Moskaluk and M.H.Stoler, Diagnostic Molecular Pathology, 2002, p234-238) set forth cultured cell is embedded in and make polyclonal cellular array in the agarose and place on the slide.Zhu (United States Patent (USP) 7,598,036) has set forth protein, nucleic acid, or cultured cell in vitro is directly on slide, and perhaps point is attached to microscope slide as external contrast with film with test sample again on special film earlier.Set forth a kind of in order to the method for the polypeptide of covalent bond on solid support as the IHC contrast.

Summary of the invention

The present invention is relevant for a kind of method based on wide spectrum quality control apparatus and production and this device of use of cellular array of carrying out quality control in immunity and molecular pathology detection.

One aspect of the present invention is the quality control cellular array.Cellular array is made of a plurality of independent zone (point) that disperses that various kinds of cell constitutes.The cell that cellular array comprises can be normal cell, also can be optimum or malignant cell.An embodiment of quality control cellular array is to comprise that origin comes from kinds of tumors or cancer type (carcinoma cancer, the melanoma melanoma, the sarcoma sarcoma, leukemia/lymphoma leukemia/lymthoma, and tumors of the neural system and nervous system neoplasm) the cell that the separates point formed of cultured cell.The mixture of a kind of cell or several cells is by o'clock in a lip-deep distinct area of a solid support, and this zone is called as the cell point.Cellular array is made up of a plurality of cell points, can place microscope slide surface or places on the film or film that can be attached to little surface of glass slide.

An embodiment of the invention are that the Quality Control cellular array comprises isolated clone from cancerous tissue.Another embodiment of the invention is that the Quality Control cellular array comprises isolated clone from lymphoma tissue.Another embodiment of the invention is that the Quality Control cellular array comprises from melanoma, sarcoma, or isolated clone in the nervous system neoplasm.

Another embodiment of the invention is that the Quality Control cellular array comprises cell infected by microbes or that express microbial gene.

Another embodiment of the invention is that each clone in the Quality Control cellular array is positive control to some detection, is negative controls to other detections simultaneously.Another embodiment of the invention is that the cell in the Quality Control cellular array is that in vitro culture obtains, and handles through chemical reagent.The chemical reagent of handling cell comprises formaldehyde, valeral, ethanol, acetone, acetic acid, dimethylbenzene, perhaps dimethylbenzene substitute.Another embodiment of the invention is that the Quality Control cellular array comprises and is less than 50 cell points.Another embodiment of the invention is to contain 102 to 104 cells in each cell point of Quality Control cellular array.Another embodiment of the invention is the mixture that each cell point of Quality Control cellular array comprises one or more cells.

Another embodiment of the invention is that the Quality Control cellular array is put on micro-slide.Described micro-slide has a marked region and a surveyed area that comprises the Quality Control cellular array.Another embodiment of the invention is that the Quality Control cellular array is put on micro-slide.Described micro-slide has a marked region, and sample areas and another one comprise the zone of Quality Control cellular array.Another embodiment is a label that comprises patient or sample information that is attached to the mark zone, the predetermined expression general layout of a certain detection target on the Quality Control cellular array that is imprinted on the label, and (selectable) detects the date.Another embodiment is that Quality Control cellular array device further comprises and determines one group by the target that is detected by the detection method of Quality Control.Another embodiment is that Quality Control cellular array device further comprises the data group that a predetermined contact detects target and Quality Control cellular array.

Another aspect of the present invention is the method for making Quality Control cellular array device.At first, determine one group of target that is carried out the detection method detection of Quality Control by Quality Control cellular array device.According to the detection target group of selecting, determine to be contained in the clone in the cellular array.Detect the expression that all should in a selecteed clone, be positive at least of each target in the target group.Alternatively, detect all members in the target group and in selected clone, have the expression that is negative of a clone at least.Detect to be qualitatively or quantitatively determined in the cell that target refers to that test sample and Quality Control cellular array comprise and existence or non-existent protein, gene, RNA molecule.Detect step and refer to SABC, immunocytochemistry, and in situ hybridization.For SABC and immunocytochemistry, detect target finger protein antigen.For in situ hybridization, detect target and refer to DNA or RNA.

An enforcement aspect of the present invention is that following detection target is included in the target group: pencytokeratins, EMA, S-100, HMB45, smooth muscle specific antigen, ER, PR, Her2, P53, PSA, vimentin, chromogranin, desmin, CD3, CD20, CD117/c-kit, CD15, CD30, CD45, CD99, Bcl-1 (cyclin D1), Bcl-2, Ki-67, Ig kappa, Ig lamda, and CEA.According to our research, more than 26 cell proteins be the most frequently used biomarker in daily SABC or the cellular immunity test clinically.They form the center detection target group that the Quality Control cellular array carries out quality control.

Another aspect of the present invention is to use the method for Quality Control cellular array device.Comprise and set up one at the predetermined data group of cellular array and relevant detection target group; The data group is presented on Jie of paper media or the electronic media; Carry out testing procedure in specimen and Quality Control cellular array with the reagent that test detects the member in the target group simultaneously; With observation, imaging, or test result is noted in digital scan; And the data in test signal and the predetermined database are contrasted.

An embodiment of the invention are that predetermined data group is to be determined by following steps: on the Quality Control device each member who detects in the target group is carried out testing procedure, to observe, imaging, or digital scan notes test result, and for each the cell point in the Quality Control cellular array to each detection target.An embodiment is that the predetermined data group of Quality Control device is to be stored among digital media such as CD or the DVD.In this case, data can or be printed on the paper in the computer monitor demonstration.Another embodiment is that predetermined data group can be rendered as form, diagram, or one group of image.

Description of drawings

Fig. 1 illustrates the sample of a quality control cellular array device.The quality control cellular array is an indicia on micro-slide 1, and (Fig. 1 a) wherein to comprise a sample areas 2, a control area 3 and a marked region.Fig. 1 b points out that the label of a printing is attached to the label area of microslide.The label 5 of this printing comprises sample ID6, and concrete analysis sample ID7 is in a demonstration sample 8 of deciding in advance and any one analysis data presented storehouse 9 of quality control cellular array concrete analysis target.Fig. 1 c explanation is in the signal indication of the test result of quality control cellular array.

Fig. 2 illustrate 9 on the quality control cellular array of microslide possible position.At a, b, c and d cellular array are positioned at the edge near microslide.The later test sample book on same slide of orientating as of these cellular arraies has stayed enough spaces.Be positioned at the central authorities of microslide at e and these cellular arraies of f.On slide e and f, do not leave sample areas.

Fig. 3 is the sample of a quality control cellular array.Ten cell points are aligned to two these cellular arraies of row and comprise five cancer cell systems, two lymphoma cell lines, a melanoma cell series, a sarcoma cell line and a cns tumor clone.

Fig. 4 is the outline demonstration of setting up a predetermined database for the quality control cellular array.Carry out an IHC or ISH analysis at objectives at the quality control cellular array by antibody or probe, be included in the detection target group and be controlled by quality cellular array control.This analysis result reads the microslide that carries the quality control cellular array by naked eyes or machine and comes record, and is stored on the paper or on the digital media.Quantitative or semiquantitative indication all is assigned on the test signal in test result.These programs need be repeated, and each evaluating objects all is included in detects in the target group, are used for setting up a quality control cellular array and the database relevant with detecting the target group.

Fig. 5 has showed at IHC, the step that service property (quality) control cellular array was analyzed during ICC or ISH analyzed.IHC, ICC or ISH analyze and carry out simultaneously in test sample book with on the quality control cellular array in test sample book, pass through antibody or probe at interested target under same condition, are included in to detect to be controlled by quality cellular array control in the target group.This analysis result, that is, this signal pattern, be created on the quality control cellular array observed and by with in advance the ruling database in information compare.The test success of this sample of coupling expression.Do not match and represent the test crash of this sample.

Fig. 6 has showed foundation with use and quality control cellular array and has detected the relevant computer software of target group and the program of database.Two parallel information flows are required.First information flow is that ID or the antibody/probe of manual typing evaluating objects enters computer software.Second information flow facilitated by machine, needs image and input target or the antibody/probe I D of painted cellular array, passes through machine by readable digital marking (for example bar code).Be these two information flows, computer software is that each test antibody/probe distributes an ID (for example numeral or bar code), and is relevant with the painted areas of specific antibodies/probe, in the database of deciding in advance of cellular array, be stored in the computer, and by analyzing decimal system software.This computer is that specific objective or antibody/probe are printed painted pattern, for manual program, namely is used for comparing with painted pattern in Realistic Analysis by surgical staff.In machine stream, the rendered image that needs is to compare by the image in the database of analysis and measurement and ruling in advance, confirms that then the result will be generated.This slide read-out device can be installed in and realize automated quality control on the automatic staining device.This slide read-out device also can be used as an automated quality control proofreading equipment.

The specific embodiment

The present invention relates to the device that quality control is provided for immunohistochemistry, cytology and molecular pathology analysis, and use this device that the detected material in the cell or tissue section is analyzed, especially during immunohistochemical test, realize the method for quality control.It needs to be noted, the present invention relates to the making quality control apparatus, comprise the method for making cellular array, and in correlation analysis, use such cellular array to realize the method for quality control.Specifically, quality control apparatus involved in the present invention comprises an a plurality of independent zone that disperses that is made of dissimilar cultured cells, and these cells are fixed and adhere to through the plane of agent treatment surface of solid phase carriers.This plane solid phase carrier can be divided into front (also can this as top) and back (also can this as the bottom surface).The detection platform that this plane solid phase carrier can be a plane (as, be used for fractographic slide), also can be that adhesive by the bottom surface adheres to the film on the detection platform.

1. quality control cellular array device

An aspect of this invention is the quality control cellular array, i.e. a plurality of independent zone (point) that disperses that is made of various kinds of cell.Mentioned herein to cell can be normal cell, also can be optimum or malignant cell.Can be directly from the tissue or body fluid isolated cells, also can be cultured cell.Staging (type) comprising: cancer, melanoma, sarcoma, lymthoma/leukaemia and nervous system knurl.The mixture of the cultured cell of single kind or two to three kinds of dissimilar cultured cells is loaded the isolated area that is separated from each other in surface of solid phase carriers, is referred to herein as the cell point.

Professional term, cancer, sarcoma, leukaemia/lymthoma, myeloma and melanoma are commonly used to distinguish the tumour that different tissues originates from.Cancer is a kind of epithelial tumor type that originates from, and is modal malignant tumour type, accounts for the 80-90% of all malignant tumour cases greatly.Sarcoma is a kind of tumor type that originates from muscle, bone, cartilage, fat or connective tissue.Leukaemia originates from white blood cell and precursor thereof.Lymthoma is the malignant tumour that originates from bone marrow derived cell, and involves lymphatic system.Leukaemia and lymthoma all are malignant tumor of hematopoiesis system.In the application's book, leukaemia and lymthoma are collectively referred to as the hemopoietic system malignant tumour.Melanoma is to originate from melanocytic malignant tumour, and melanocyte can be produced as skin, hair, the painted melanin of eyes.

(Fig. 1 is special microslide 1 a) for one of concrete scheme of quality control apparatus, it has by a plurality of and is dispersed in the quality control district that distribution, separate cell point 3 constitute, each cell point is formed by individual layer or near the cultured cell of individual layer, and the mixture of every kind of cell or two to three kinds of cells is restricted in the individual cells point.Generally speaking, cell adheres to carrier surface by covalent bond.When tissue need be attached at the slide of electric charge processing, cell also can adhere to carrier surface by charge effect.This slide also comprises a sample areas 2 and a label area 4.Two of the concrete scheme of quality control apparatus, the label of slide (Fig. 1 b), it comprises a sample identification marking 6, a target (or antibody/probe) identification marking 7, also be printed on the expression pattern 8 of predetermined particular target in cellular array on the label, this writes the routine analyzer of specific expression pattern by the Quality Control cellular array also can show 9, can also selected marker the zone of date of test.Three of the concrete scheme of quality control apparatus provides a series of and pre-determines, the diagram of the expression pattern in cellular array 8 of various targets (Fig. 1 c).This diagram comprises, but be not limited to following three: the solid dot that (A) is filled with different uneven colors, be example with Fig. 1 cA, the lightest point represents background dyeing (maybe can not detect target/analyte), along with the intensification of solid dot fill color, represent critical expression (+/-) step by step, the weak positive (+), middle positive (++), and strong positive (+++); (B) letter that has a circle is used to indicate the Subcellular Localization of the target that detects, be example with Fig. 1 cB, hollow circle is designated as background signal (feminine gender), there is the alphabetical n of circle to be designated as positive signal and is positioned at nucleus, there is the alphabetical c of circle to be designated as positive signal and is positioned kytoplasm, have the m of circle letter to be designated as positive signal and be positioned cell membrane; (C) circle of Bian Huaing, be example with chart 1cC, the circle that dotted line surrounds is designated as background signal (feminine gender), the adularescent point is designated as the cytoplasm signal in the circle that the solid black lines surround, have the black point to be designated as the nucleus signal in the white circle, the circle that is with a circle heavy black line bar is designated as the cell membrane signal.

Four of the concrete scheme of quality control apparatus, surface adhesion has the film of a plurality of independent cell points, and the bottom surface of this film can adhere to the test surfaces of selected detection platform, for example, microslide.

Cell preferably is individual layer and adheres to carrier surface, but because of the agglomerating growth of cell, also can form multi-layer cellular.Cultured cell can be placed on carrier surface before fixing and processing.At this moment, cell need be fixed and handles at solid phase carrier.But had better at point sample before solid phase carrier, by chemical reagent cell be fixed.Treatment step generally includes, dehydration of alcohol, dimethylbenzene or transparent, the waxdip/embedding of dimethylbenzene substitute.These steps are similar to the handling procedure of tissue samples.In the present invention, prepare the used treatment step of cellular array and can simplify, save dehydration and transparent step.

Five of the concrete scheme of quality control apparatus, polytype cell are embedded in and make the wax stone that includes the various kinds of cell type in the paraffin.These cell wax stones can further be made into section, and adhere to surface of solid phase carriers.A kind of method of making various kinds of cell type wax stone is that cell fixing and that handled is made into the columniform core of a branch of bundle, and they are embedded in the paraffin; Another kind method is, fixing and handle the cell of particular types in the capillary that can be cut off by microtome, a plurality of such capillaries are assembled bunchys, and are embedded in the paraffin.The quality control cellular array can be placed on same surface of solid phase carriers with sample or be positioned on the independent carrier.This carrier can be microslide, transparent film, film or the cover glass that can adhere to quality control and/or detect sample.Transparent film or film can be made with glass as nylon, polystyrene, polypropylene, cellulose nitrate etc. by various macromolecular materials.Film or film be the solid phase plane of 0.001mm to 0.5mm thickness preferably.

Six of the concrete scheme of quality control apparatus, the cell in each independent point can be used as the positive control of some detection, can be used as the negative control of other detection simultaneously again.Quality control cellular array among the present invention is a low-density array, and the independent cell point that it comprises is no more than 50, and better situation is to be no more than 20 cell points, be less than 12 then better.The few reason of the quantity of the cell point that cellular array in the present invention comprises is the manufacturing of being convenient to the cellular array device, makes things convenient for observation and the analysis of testing result simultaneously.Cell quantity in this invention in each cell point is between 10 to 105, and is best between 102 to 104.

2. select to be used for the clone of quality control cellular array

One of concrete scheme is identified for clone in the quality control cellular array by the detection to some row targets.Preferred plan is, forms cellular array by the least possible cell point, can carry out quality control to clinical correlation analysis as much as possible again simultaneously.Here, analysis refers to detect specific measured object, target or biomarker.For example, detecting a kind of target molecule albumen with a kind of antibody by immunohistochemical method is an analysis, and detecting another kind of target molecule albumen with the method for another kind of antibody by the SABC chemistry is exactly the another one analysis.

A kind of clone refers to single origin, purified cultured cell group.Between nearly decades, massive tumor clone is established, and these clones come from the malignant tumor tissue of nearly all type and origin.About 1000 kinds of clones that come from human malignancies have been included in the ATCC catalogue.The clone that derives from tumor tissues or cancerous tissue is many cultivates by going down to posterity or virus transfection obtains, so they can express the antigen that great majority are expressed in primary tumo(u)r or cancer.By standardized cultural method, be easy to obtain the cultured cell of a large amount of different clones.

Cultured cell comprises thousands of kinds of different proteantigens and other biomolecule.Yet, in thousands of kinds of different protein and biomolecule, have only minority that clinical diagnosis and prognosis are judged to have value.These molecules are commonly called biomarker.In a lot of cultured cells, the expression way of biomarker is similar to its expression in the pathological tissues cell.The clone that has potential using value in the present invention derives from multiple malignant tumour, and therefore, the expression of its clinical relevant biomarkers or disappearance are similar to the expression of tumour cell in these malignant tumours.

Two of concrete scheme, the quality control cellular array can provide the target of control test to comprise a series of biomarkers with very high clinical reference value.Through identifying, there are 26 kinds of biomarkers to have very high clinical reference value, comprise keratin (pancytokeratins), EMA, S-100, HMB45, smooth muscle specific antigen (smooth muscle specific antigen), and ERs (estrogen receptor, ER), progesterone receptor (progesterone receptor, PR), Her2, P53, and PSA (prostate specific antigen, PSA), vimentin (vimentin), chromograin (chromogranin), desmin (desmin), CD3, CD20, CD117/c-kit, CD15, CD30, CD45, CD99, Bcl-1 (cyclin D1), Bcl-2, Ki-67, Ig kappa, Iglamda, and CEA.According to research of the present invention, above-mentioned 26 kinds of cell proteins are the most frequently used clinical marker things of daily immunohistochemistry or immunocytochemistry test.They have formed the core that the quality control cellular array can provide the target of control test.

Three of concrete scheme, the quality control cellular array can provide the target of control test to expand, and contains following cell protein: high molecular weight cell keratin (cytokeratin, HMW), and the low-molecular-weight cytokeratin (cytokeratin, LMW), cytokeratin 7 (cytokeratin 7), cytokeratin 20 (cytokeratin 20), EMA, S-100, HM 45, MART-1, tyrosinase (tyrosinase), PLAP, liver cell specific antigen (hepatocyte specific Ag), flesh specificity actin (muscle specific actin), smooth muscle specific antigen (smooth muscle specific antigen), ER, PR, Her2, P53, PSA, PSAP, flesh forms albumen (myogenin), chromograin (chromogranin), synaptic vesicle albumen (synaptophysin), desmin (desmin), vimentin (vimentin), CD99, CD3, CD7, CD10, CD4, CD8, CD20, CD117/c-kit, CD5, CD15, CD 21, CD23, CD30, CD45, Bcl-1 (cyclin D1), Bcl-2, Ki-67, Ig kappa, Ig lamda, terminal deoxynucleotidyl transferase (terminal deoxynucleotidyl transferase, TdT), calcium (looking) nethike embrane albumen (calretinin), WT-1, TTF-1, and CEA.

Four of concrete scheme, the quality control cellular array can provide the target of control test to further expand, contain following cell protein: high molecular weight cell keratin (cytokeratin, HMW), low-molecular-weight cytokeratin (cytokeratin, LMW), cytokeratin 5 (cytokeratin 5), cytokeratin 6 (cytokeratin 6), cytokeratin 7 (cytokeratin 7), cytokeratin 8 (cytokeratin 8), cytokeratin 14 (cytokeratin 14), cytokeratin 18 (cytokeratin 18), cytokeratin 19 (cytokeratin 19), cytokeratin 20 (cytokeratin 20), EMA, inhibin (inhibin), mammary gland globin (mamaglobin), Ki-67, P27, P53, terminal deoxynucleotidyl transferase (terminal deoxynucleotidyl transferase, TdT), the phosphatase of anti-tartaic acid the (tartrate resistant acid phosphatase, TRACP 5b), TTF 1, PLAP, AFP, hCG, and ERs (estrogen receptor, ER), Her2, EGFR, and progesterone receptor (progesterone receptor, PR), Zap-70, P63, and PSA (prostate specific antigen, PSA), prostate specific acid phosphatase (prostate specific acid phosphatase, PSAP), liver cell specific antigen (hepatocyte specific Ag), chromograin (chromogranin), synaptic vesicle albumen (synaptophysin), CD1a, CD3, CD4, CD5, CD7, CD8, CD10, CD15, CD20, CD21, CD23, CD30, CD34, CD35, CD43, CD56, CD57, CD79a, CD99, CD117/c-kit, Bcl1 (cyclinD1), Bcl2, Bcl-6, between become lymthoma kinases-1 (anaplastic lymphoma kinase-1, Alk-1), fascin, Ig Kappa, Ig Lamda, Pax-5, P 504s, MART-1, HMB45, S-100, PAP, NF, tyrosinase (tyrosinase), calcium (looking) nethike embrane egg (calretinin), WT-1, CEA, Cox-2, flesh specificity actin (muscle specific actin), smooth muscle actin (smooth muscle actin), desmin (desmin), flesh forms albumen (myogenin), and vimentin (vimentin).

Five of concrete scheme, the quality control cellular array can provide the target of control test to further expand, contain following cell protein and biomarker: flesh specificity actin (muscle specific actin), smooth muscle actin (smooth muscle actin), Alk-1, Bcl-1, Bcl-2, Bcl-6, BF-1, CD1a, CD3, CD4, CD5, CD7, CD8, CD10, CD15, CD20, CD21, CD22, CD23, CD25, CD30, CD31, CD34, CD35, CD38, CD43, CD44, CD45, CD56, CD57, CD68, CD79a, CD117/C-kit, CD138, CEA, chromograin (chromogranin), TRPM-2 (clusterin), the high molecular weight cell keratin (cytokeratin, HMW), low-molecular-weight cytokeratin (cytokeratin, LMW), cytokeratin 5 (cytokeratin 5), cytokeratin 6 (cytokeratin 6), cytokeratin 7 (cytokeratin 7), cytokeratin 8 (cytokeratin 8), cytokeratin 14 (cytokeratin 14), cytokeratin 18 (cytokeratin 18), cytokeratin 19 (cytokeratin 19), cytokeratin 20 (cytokeratin 20), desmin (desmin), EMA, fasciclin (fascin), granzyme B (granzyme B), HBME, hemoglobin (hemoglobin), IgA, IgD, IgE, IgG, IgM, J-chain, Ki-67, Ig kappa, Ig lambda, LMP-1, MUM1, myeloperoxidase (myeloperoxidase), NSE, Ab OCT-2, S-100, Pax5, TDT, TIA-1 TRAP, TTF-1, Vs38c, vimentin (vimentin), Zap-70, CD83, P62, CK18, CD163, P63, P16, Stat-3, calcium (looking) nethike embrane egg (calretinin), BerEp4, CK5/6, ubiquitin (ubiquitin), tyrosinase (tyrosinase), PR, ER, synaptic vesicle albumen (synaptophysin), E-cadherin (E-cadherin), MyoD1, ommatidium associated transcription factor antibody (microphthalmia), MUC-1, MUC-2, laminin (laminin), flesh forms albumen (myogenin), HER2, EGFR, MOC-31, CD99, TAG-72, MLH-1, MSH-2, AR, villin (villin), melanic related antigen (melanoma assoc Ag), ACT, alpha-1 antitrypsin (alpha-1-antitrysin), P27, P504s, inhibin (inhibin), PSA, AFP, MUC-5, MUC-6, P21, fibronectin (fibronectin), P53, beta-catenin (beta-catenin), CD123, CD24, CD133, adenovirus antigen (adenovirus antigens), cytomegalovirus antigen (cytomegalovirus (CMV) antigens), Epstein-Barr virus antigen (Epstein-Barr virus (EBV) antigens), Heliobacter pylori antigen (Helicobactor pylori antigens), HPV antigen (human papillomaviruses (HPV) antigens), HBcAg, HSV I﹠amp; II antigens, Her2 gene magnification (Her2 gene amplification), c-myc gene dystopy (c-myc gene translocation), poly-A mRNA, Ig kappa mRNA and Ig lambda mRNA.

Six of concrete scheme detects the target group and can include only one or more microbial antigens or gene.In the case, the cell in the cell point that comprises of quality control cellular array comprises antigen or the gene of one or more microorganisms.Simultaneously, the quality control cellular array also can comprise a cell point that does not contain microbial antigen or gene, as negative control.

Only 10 cell points detect the above-mentioned several groups of needs (Fig. 3) that the target of expanding carries out quality control with regard to being enough to satisfy to immunohistochemical method, for each target, have at least a cell point to be positive.Cellular array shown in Figure 3 comprises 5 points that come from malignant cell system, and 2 individual from lymphoma cell line, and the clone of melanoma, sarcoma and neurogenic malignant tumour respectively has 1 point.

Except the related albumen target of immunohistochemistry IHC, the nucleic acid target that the quality control cellular array also can be among the cross experiment ISH in position provides contrast.As, c-myc gene dystopy, Her2 gene magnification, poly-ARNA and kappa/lambda mRNA are exactly the contrast of in situ hybridization test ISH.

In immunohistochemistry IHC, detecting the target group is proteinaceous antigen, can detect by specific antibody.In the present invention, detect the target group and can comprise all above-mentioned marks, in addition more, as, the biomarker of the useful clinical value of new discovery and introducing.In this case, the quality control cellular array must be able to comprise the cultured cell system of greater number.Judge or the research needs according to specific diagnosis, prognosis, can work out the detection target group of reduction targetedly.At this moment, make up the cellular array needs less relatively clone only just meet the Quality Control requirement.

Be to realize the quality control to selected specific detection target group, each composition is should be at least expressed and can be detected by a kind of clone in the constructed cellular array in the group, so that positive control to be provided; Each composition is should be at least not expressed and can not be detected by a kind of clone in the constructed cellular array yet in the group, so that negative control to be provided.Yet if detect detection target or the label that includes wide expression in the target group, cellular array just can not provide negative control for this target.Detection target or the label of wide expression, as: PCNA, GAPDH, albumen (house-keeping gene GAP-associated protein GAP) of beta-actin and much basic physiological function metabolism adjustings of other participation cell can be expressed in all cells system.The concrete scheme that addresses this problem is, adds a non-humanized's clone in cellular array, as the negative control of the human biomarker of wide expression.The concrete scheme that clone is selected in this invention comprises that also in the quality control cellular array, the detection target for a specific non-wide expression is no more than 75%, 50%, 25%, or 10% cell point can produce the positive signal that can be detected.This invention also is embodied in, and in detecting the target group, the target of wide expression indicated, and is different from other targets in the group.The signal strength signal intensity that noun " detectable level " refer to produces in detection can show expression or the existence of a certain target or label, can be color or fluorescence signal; " detectable level " refer to be higher than background noise signal strength signal intensity." undetectable level " refer to can not with other signal strength signal intensity of background noise phase region.

A concrete scheme of this invention is that the cell point that a non-humanized's of adding clone constitutes in the quality control cellular array comes to be all targets, comprises the target of wide expression, and negative control is provided.Here, non-humanized's clone is not limited to certain specific non-Humanized cell system.The specialty the researcher think, according to the needs of particular detection target group, from multiple organism as: the clone of mouse, ox and insect all can be used for cellular array of the present invention.

Detect the target group and also can comprise from microprotein and nucleotide sequence, as: virus, bacterium, yeast and fungi.If these targets are involved, just can provide Quality Control for detecting the cell infection and the conversion that are caused by related microorganisms by the quality control cellular array.

Detect the target (albumen or gene order) that the target group can an origin comes from microorganism and form, also can both comprise the microorganism target, comprise the target that derives from cell again.The target that derives from microorganism can also can import microbial gene or genetic fragment and be expressed in the cell by introducing in the cell point with microorganism direct infection cultured cell by engineered method.

3. the using method of quality control cellular array

This invention also comprises device using method as described herein.Cellular array Quality Control slide can use separately, or the test zone of point on slide, and its test zone can load sample to be tested, and (Fig. 1 a).Confirm and verify the reliable method of result of the test of immunohistochemistry IHC, immunocytochemistry ICC, in situ hybridization ISH, comprise whether the target that detects in the biological specimen exists and celluar localization.Method comprises, when detecting some or a plurality of target molecules, handles the cellular array quality control apparatus among biological sample and the present invention synchronously.Here used noun " processing " refers to carry out to detect whether target molecules exists and all programs of Subcellular Localization.As, " processing " can be the program of carrying out the IHC of immunohistochemical test, by detecting the existence of target protein with the antibody of this albumen specific bond.Under appropriate condition, antibody is combined with the target protein specificity, detect the signal (as: chrominance signal of chromogenic reaction generation) that the antibody be combined with target protein carries again, detect the existence that these signals can be indicated target protein, can not detect the signal instruction target protein and not express.The researcher of specialty is familiar with more such test procedure very much.In part for example, we provide standard immunohistochemistry IHC and in situ hybridization ISH operating process.

This device can also be required to be cellular array and set up upright preset database with relevant detection target by correct use.Preset database will store with the form of paper material or digital media and provide.For setting up presetting database, on Quality Control cellular array device, need test in a large number, with related reagent each component that detects in the target group is independently tested.For example, for setting up immunohistochemical test's database that 26 molecule cores detect the target group, will be on cellular array device independently, by specific antibody, according to the operating process that provides in the example 2, each component is wherein carried out independent experiment.All cells in the Quality Control cellular array all will react with detection reagent.For specific target, the some or all cells in some cell point will produce positive signal, and all cells will produce negative signal in other cell point.

Noun " positive signal " refer to is higher than background signal at the signal of the detection reagent place generation of interested particular detection target, when perusal or machine detection, is easy to and the differentiation of nonspecific background signal.Noun " negative signal " refer to, can not be distinguished with nonspecific background signal when perusal or machine detection at the signal of the detection reagent place generation of interested particular detection target.These signals will be recorded and analyze, and provide indication digital or numeral.For example, the strong positive signal can change high pixel into, or is designated as a plurality of "+" symbol.For example, " +++" the expression epistasis, positive in " ++ " expression, the weak positive of "+" expression, "+/-" represent the uncertain positive, "-" expression is negative.Except the signal strength signal intensity of the target expression in the reacting cells, in analytic process, can also show and record the Subcellular Localization that detects each component in the target group.After the information of quantitative or semi-quantitative results and Subcellular Localization that acquisition detection target group all components is expressed in all cells point of quality control cellular array, we have just set up required presetting database.Database can store by paper material, comprises but is not only: catalogue, chart or pictures.Database also can be stored and presents by a digital media, for example computer monitor and CD/DVD.

As detect the target that comprises in the target group and in the cell point of quality control cellular array, form typical positive signal, can be used as individual authentication, prove that the operation that detects target in biological sample is accurate.When biological sample produced negative findings, as non-coloring or fluorescence signal, it is particularly important that such contrast seems.The positive findings that the quality control cellular array produces can confirm that The above results is real negative findings, but not the problem of the mistake of program or reagent quality causes.Because biological specimen and quality control cellular array are adhered on same the microslide, and under same condition, contact with same reagent, therefore form checking reliably.

Another concrete scheme comprises, determines the analytical method of target molecule Subcellular Localization in biological specimen.Method comprises, in immunohistochemistry IHC or in situ hybridization test ISH, as mentioned above, handles biological specimen and quality control cellular array simultaneously.Result shows as target molecule specific subcellular area in the cell of the cell of biological specimen and quality control cellular array and exists or do not exist.The Subcellular Localization of the signal that produces compares in the Subcellular Localization of the detectable signal that target molecule produces in the biological specimen and its cell point in the quality control cellular array.

Subcellular location includes, but are not limited to: nucleus, cytoplasm, cell membrane and nuclear membrane.The expression of particular target and their Subcellular Localization are constant in the specific cells system of quality control cellular array, namely are determined and record in design and making quality control cellular array.Biological sample and quality control cellular array are carried out a certain test procedure, detect certain target molecules the result can with pre-determine and record, Subcellular Localization and the expression of this molecule in cell compares in the quality control cellular array.As the data consistent of the result of current test in the quality control cellular array with certain particular target that pre-determines and record, prove that the operation of current test is successful, test the credible result in biological specimen specifically, can send report.Inconsistent with the data of certain particular target that pre-determines and record as the result of current test in the quality control cellular array, then the operation of the current test of explanation has problems, and the result in biological specimen is insincere in this test, can not send report.

The present invention also comprises a commercial kit.Kit is by quality control cellular array device, can realize one group of quality control information that detects target by array, what pre-determine and record comprises in the quality control cellular array expression and the Subcellular Localization database of information of each detection target in each cell point.The expression of target can come the quantity of quantitative or "+" symbol to come sxemiquantitative by digital pixel.For example, " +++" represents strong positive, and " ++ " represents positive, and "+" representative is weak positive, and "+/-" representative is uncertain, and "-" represent feminine gender.Wherein also include one and recorded each and detect target at form or the computer file of the expression of each point, and print pass through scanning each detection target that obtain or digital version in the image result of the expression of each point.

Selection and controlled detection target group that example 1 is included in the clone in the quality control cellular array have close relation.In this example, 11 clones are selected the quality control cellular array that is made into 11 points, comprise clone Hela (adenocarcinoma of the uterine cervix), LS174T (colorectum gland cancer), SW480 (gland cancer, colorectal), BT474 (breast duct carcinoma), MCF-7 (breast cancer), 22Rv1 (prostate cancer), Ragi (B cell, Burkitt lymphoma), HuT 78 (t cell lymphoma), Jurkat (T chronic myeloid leukemia), SK-UT-1 (sarcoma), SW1353 (sarcoma), and CaC1 (melanoma).These 11 kinds of clones of selecting from the quality control cellular array, the core group that contains 26 members of target protein, to a certain extent, each target protein is in this core group, all express a detectable level at least one clone, target protein shows the level that can not realize at least one clone simultaneously.

Example 2 immunohistochemistries (IHC)

In this example, a biological specimen (comprising the quality control cellular array) is by antibody treatment (primary with less important), and that had handled gives the chromogen color development, gives counterstain then at last.

A. deparaffinization

When the histotomy that uses paraffin to implant, deparaffinization must be performed, as representational be to soak through 52 minutes of taking turns dimethylbenzene, twice 100% alcohol, one time 95% alcohol makes rehydration, air mummification at room temperature is five minutes then.

B. antigen obtains

Using the skill of specific antibody well-known in IHC is painted, is to obtain program preliminary treatment formalin set histotomy by an antigen, normally obtains in the solution at antigen and handles by heating or proteolytic enzyme.After antigen obtained, sample comprised the quality control cellular array, was used to contain 5% the normal sheep of mixing or horse serum and hatched at room temperature 20-30 minute.

C. initial and antigen-reactive secondary

And then by the hatching of serum, sample is washed with PBS.Prediluted initial antigen serum or antibody (approximately 150-200mu.L) are applied to each sample.Sample was at room temperature hatched 2-4 hour by antigen serum or antibody, or hatching 2 hours in 40 degrees centigrade environment of a humidity, perhaps can hatch a whole night in the environment of the humidity of a room temperature.At the sample on slide after the hatching, will in staining dish, wash 3 times by PBS.

The secondary antibody that waters down (approximately 150-200mu.L) is used in each sample on slide.This was by hatching in the environment of one 40 degrees centigrade humidity 30 minutes.Sample after hatching will wash in staining dish 3 times by PBS.

D. give birth to removing of peroxidase activity in

Sample on slide then is placed in the 500mlPBS solution that contains 3% hydrogen peroxide and 1% sodium azide, and at room temperature hatches and give birth to peroxidase activity in removing in 15 minutes.By the sample on slide after the hydrogen peroxide PBS hatching, will in staining dish, wash 3 times by PBS.

E. color development

(Vector Laboratories Inc., Burlingame is Calif.) by using their active set of PBS dilution for the ABC synthetic.This active set (approximately 150-200mu.L) is applicable to each sample on slide.The hatching of ABC solution is in the environment of one 40 degrees centigrade humidity 30 minutes.The sample on slide after the hatching will wash in staining dish 3 times by PBS.

DAB solution is to prepare with the solution that interpolation 50.mu.L contains 30%H2O2 by adding 100mg DAB to 100mL PBS.Approximately the DAB solution of 150-200.mu.L is added in each sample on slide.Color development can monitor by microscopic examination.Coloured precipitation will form at positive cell.Color began to manifest after 2-5 minute, reached enough imagings usually in 10 minutes, but 20-30 minute hatching may need to be used to softer weak colo(u)r atlas.In order to stop development, will in staining dish, wash 3 times by deionized water at the sample on the slide.

F. painted on the contrary

Sample on slide immerses 10-50 second in the haematine of Harris, and cleans for three times by being dipped into deionized water, is immersed in then in 0.2% Ammonia 30 seconds, and cleans for three times by being dipped into deionized water.They immerse in 95% ethanol twice, and each 2 minutes, next immerse in 100% ethanol twice, each 2 minutes, last, this slide cleaned by immersing in the dimethylbenzene twice in each 2 minutes.

Example 3 in situ hybridizations

In this example, biological specimen homogenous quantities control cellular array (also being used as sample from now on quotes) is on slide, by vitamin or digoxigenin label probe, with toxamin or anteiso-gitoxigenin antibody generation chemical reaction.Therefore this sample is colored.

A. deparaffinization

Sample on slide is by putting into dimethylbenzene four times, each 5 minutes, in 100% ethanol twice, in each 1 minute and 95% ethanol twice, comes deparaffinization in each 1 minute.(Calif.) deionized water is cleaned and washes the histotomy slide of deparaffinization for BioGenex, San Ramon by containing RNase Block then.

B. the Proteinase K of mounted tissue samples is handled

Approximately the fresh Proteinase K solution that waters down of 150-200.mu.L is placed on each on the sample of slide, and at room temperature hatches 15 minutes.After the digestion, the sample on slide was washed 5 minutes at staining dish by RNase Block by the PBS of 500mL.Following solution dewaters sample by immersing in the staining dish in succession: 500mL distilled water adds RNase Block10 second, the ethanol of 500mL 50% adds RNase Block10 second, 10 seconds of ethanol of 95% of 500mL and 10 seconds of 500mL 100% ethanol.Whole slide at room temperature dry 5 minutes.

C. with the make marks hybridization of probe of vitamin or digoxigenin

The hybridization solution that comprises vitamin or digoxigenin labeled oligonucleotide probe is placed on the sample of each slide.This needs the solution of about 50-100.mu.L.Slide be placed in the baking box or heat block on, with 95 degrees centigrade 8-10 minute, change the habit of nucleic acid.This step has been eliminated the hairpin loop of mRNA order or has been turned over glue.After the step that changes one's skin, slide is hatched a whole night by under 45 degrees centigrade in a wet environment.Follow hybridization step, slide is cleaned 2 times in staining dish.SSC (standardized citrate solution) 37 degrees centigrade following 5 minutes, next by at 1 times SSC 37 degrees centigrade of flushings in following 5 minutes.Next this washed 30 minutes down at 60 degrees centigrade by the SSC at 0.2 times.At last, this slide was by PBS flushing 2 times, each 2-5 minute.

D. signal detection

Slide is contained the 5% hybrid standard goat of 500mL and the serum of horse, at room temperature 20 minutes by the vertical staining dish of putting into.The mouse toxamin that waters down in advance or mouse anteiso-gitoxigenin antibody (150-200.mu.L) are applied to each on the sample of slide.This contains hatching 2 hours at the sample on the slide of antibody under the environment of 40 degrees centigrade humidity.After the hatching of toxamin or anteiso-gitoxigenin antibody was arranged, this was cleaned 3 times by PBS on staining dish at the sample on the slide.

E. the application of secondary antibodies

The secondary antibody that waters down in advance (approximately 150-200.mu.L) is used in each in the sample of slide, and hatching 30 minutes in the environment of 40 degrees centigrade humidity.After the hatching, the sample on slide is cleaned 3 times at painted dish by PBS.

F. give birth to removing of peroxidase activity in

Sample on slide is placed in the painted dish that the PBS that into contains 500mL contains 3% hydrogen peroxide and 0.1% sodium azide, and at room temperature hatches 15 minutes.By the sample on slide after the hydrogen peroxide PBS hatching, washed 3 times at painted dish by PBS.

The application of G.ABC synthetic " ELITE "

The ABC synthetic is by using their active set of PBS dilution.This active set (approximately 150-200mu.L) is applicable to each sample on slide, and hatching 30 minutes in the environment of one 40 degrees centigrade humidity.The sample on slide after the hatching will wash in staining dish 3 times by PBS.

H. use the chromogen color development of diaminobenzidine tetrahydrochloride (DAB)

DAB solution by add among 100mg DAB to the 100mL PBS and add 50.mu.L contain that 30% H2O2 prepares.Approximately the DAB solution of 150-200.mu.L is added in each sample on slide.Color development can monitor by the sample that DAB observes on slide at microscopically.Coloured precipitation will form at positive cell.Color began to manifest after 2-5 minute, reached enough imagings usually in 10 minutes, but 20-30 minute hatching may need to be used to softer weak colo(u)r atlas.In order to stop development, will in staining dish, wash 3 times by deionized water at the sample on the slide.

I. counterstain

Sample on slide immerses 10-50 second in the haematine of Harris, and cleans for three times by being dipped into deionized water.Slide was immersed in 0.2% Ammonia 30 seconds then, and cleaned for three times by being dipped into deionized water.They immerse in 95% ethanol twice, and each 2 minutes, next immerse in 100% ethanol twice, each 2 minutes, last, this slide cleaned by immersing in the dimethylbenzene twice in each 2 minutes.

Claims

1. the quality control apparatus that pathology detect is characterized in that comprise cellular array, described cellular array is characterised in that and comprises multiple different cell; Wherein said cell is attached to the surface of a solid substrate; Simultaneously, wherein said various cell is arranged to many isolated cells points.

2. according to the device in the claim 1, it is characterized in that described cellular array comprises the cell line that is derived from cancerous tissue.

3. according to the device in the claim 1, it is characterized in that described cellular array further comprises the cell line that is derived from sarcoma.

4. according to the device in the claim 1, it is characterized in that described cellular array further comprises the cell line that is derived from nervous system neoplasm.

5. according to the device in the claim 1, it is characterized in that described cellular array comprises the cultured cell that contains microbial antigen or gene.

6. according to the device in the claim 1, it is characterized in that described cellular array further comprises the cell line that is derived from pernicious hematopoietic tissue.

7. according to the device in the claim 1, it is characterized in that, described cell be in vitro culture and handled with chemical reagent, used chemical reagent is select from formaldehyde, acetaldehyde, alcohol, acetone, acetic acid, dimethylbenzene, dimethylbenzene substitute and paraffin.

8. according to the device in the claim 1, it is characterized in that the quantity of the cell point of mentioning in the described cellular array is between 1-50.

9. according to the device in the claim 1, it is characterized in that the quantity of the cell point of mentioning in the described cellular array is between 5-50.

10. according to the device in the claim 1, it is characterized in that the cell quantity in the described cell point is between 100-10000.

11. the device according in the claim 1 is characterized in that, wherein said cell point comprises that a pure specific cultured cell or one are by the specific cell mixing of variety classes

12. the device according in the claim 1 is characterized in that, preferably comprises the cell of being crossed by infected by microbes.

13. the device according in the claim 1 is characterized in that, further comprises to demarcate one group of detection target.

14. according to the device in the claim 13, it is characterized in that described one group of detection target is characterised in that and comprises following member: keratin (pencytokeratins), EMA (EMA), acid calbindin (S-100), melanoma-associated antigen (HMB45), smooth muscle specific antigen (smooth muscle specific actin), ERs (ER), progesterone receptor (PR), Her2, P53, PSA (PSA), vimentin (vimentin), chromograin (chromogranin), desmin (desmin), differentiation group CD3 (Cluster ofdifferentiation 3), CD20, CD117/c-kit, CD15, CD30, CD45, CD99, B cell lymphoma albumen-1Bcl-1 (cyclin D1), B cell lymphoma albumen-2Bcl-2, NA Ki-67, immunoglobulin (Ig) Ig (Immunoglobulin) kappa, immunoglobulin (Ig) Ig (Immunoglobulin) lamda, and carcinomebryonic antigen (CEA).

15. according to the device in the claim 14, it is characterized in that wherein said one group of detection target comprises following compositions: cytokeratin 7, cytokeratin 20, melanoma-associated antigen (MART1), tyrosinase, P-ALP (PLAP), liver cell special efficacy antigen, muscle special efficacy actin, prostate specific acid phosphatase (PSAP), myosinogen, synaptophysin, CD7, CD10, CD4, CD8, CD117/c-kit, CD21, CD23, terminal deoxynucleotidyl transferase (TdT), calretinin, the nephroblastoma (WT) and thyroid gland transcription factor (TTF-1).

16. according to the device in the claim 15, it is characterized in that wherein said one group of detection target more is to comprise following compositions: cytokeratin 5, cytokeratin 6, cytokeratin 8, cytokeratin 14, cytokeratin 18, cytokeratin 19, inhibin, mammary gland globin (mamaglobin), P27 albumen, terminal deoxynucleotidyl transferase (terminal deoxynucleotidyl transferase) TdT, anti-tartaic acid phosphatase tartrate resistant acod phosphatase (TRACP 5b), alpha-fetoglobulin (AFP), human chorionic gonadotropin (hCG), epithelial growth factor receptor (EGFR), zeta chain GAP-associated protein GAP-70(Zap-70), P63 albumen, differentiation group CD1a, CD34, CD35, CD43, CD56, CD57, DC79a, B cell lymphoma albumen-6(Bcl-6), degeneration lymthoma kinases anaplastic lymphoma kinase-1 (Alk-1), actin (fascin), B clone idiosyncratic transcription factor Pax-5, P504s albumen, positive Cycloxygenase-2(Cox-2), adenovirus, cytomegalovirus (CMV), Epstein-Barr virus (EBV), pylorus bacteroides antigen (Helicobactor pylori Ags), human papilloma virus humam papillomavirus (HPV) Ags, hepatitis B NA (HBcAg), herpes virus hominis I type and II type (HSVI ﹠amp; II Ags), transcription factor (c-myc gene translocation), polyadenous purine tail messenger RNA (poly A mRNAs), immunoglobulin (Ig) κ messenger RNA (Ig kappa mRNA) and immunoglobulin (Ig) λ messenger RNA (Ig lambda mRNA) in breast cancer Her2 GFP amplification (Her2gene amplification), the nuclear.

17. the device according in the claim 13 is characterized in that, described one group of detection target comprises microbial antigen or gene.

18. the device according in the claim 17 is characterized in that, described microbial antigen or gene come from bacterium, fungi, or virus.

19. the device according in the claim 13 is characterized in that, further comprises a database, described database is decided according to the following steps:

I. carry out a detection step with detecting one group of test reaction thing that detects a member in the target at described device,

Ii. by observation, imaging or digital scan log,

Iii. corresponding to described one group of member who detects in the target, be the cell point in each described cellular array, specify a detection signal to indicate according to result of the test,

Iv. to one group of each member who detects in the target of tell, repeat i to iii step.

20. the device according in the claim 19 is characterized in that, described detection step comprises the immunohistochemistry detection, immunocytochemistry detects and in situ hybridization detects; Wherein said detection signal comprises negative positive signal, strength signal and Subcellular Localization signal; Wherein said test signal indicates and comprises that letter indicates and shape indicates.

21. the device according in the claim 1 is characterized in that, described solid substrate comprises a mark zone.

22. the device according in the claim 21 is characterized in that, further comprises a printed label, is used for being attached to described mark zone, described printed label characteristics is to comprise the signal mode of a predetermined described cellular array.

23. the device according in the claim 1 is characterized in that, wherein said solid substrate is slide, cover glass, transparent plastic film, transparent polymer film, or the clear glass film.

24. the device according in the claim 23 is characterized in that, above wherein said solid substrate has one and below one, the attached work of wherein said cellular array is on described solid substrate, simultaneously, post adhesive below the described solid substrate, so that attached work is on other substrate.

25. a method of making the device in the claim 1 is characterised in that:

A. select one group of detection target that in a pathological examination, needs quality control,

B. select to be included in all cells strain in the described cellular array, wherein each described detection target all exists at the cell line of at least one selection, and can be detected the positive,

C. at the described cell line of in vitro culture,

D. gather in the crops the cell line in vitro culture,

E. with the cell of the in vitro culture of results with cellular array form point extremely on the described solid substrate.

26. according to the method in the claim 25, it is characterized in that wherein said one group is detected that target to be comprised: keratin (pencytokeratins), EMA (EMA), acid calbindin (S-100), melanoma-associated antigen (HMB45), smooth muscle specific antigen (smooth muscle specific actin), ERs (ER), progesterone receptor (PR), Her2, P53, PSA (PSA), vimentin (vimentin), chromograin (chromogranin), desmin (desmin), differentiation group CD3, CD20, CD117/c-kit, CD15, CD30, CD45, CD 99, B cell lymphoma albumen-1Bcl-1 (cyclin D1), B cell lymphoma albumen-2Bcl-2, NA Ki-67, immunoglobulin (Ig) Ig (Immunoglobulin) kappa, immunoglobulin (Ig) Ig (Immunoglobulin) lamda, and carcinomebryonic antigen (CEA).

27. according to the method in the claim 26, it is characterized in that, wherein said one group of detection target more comprises: cytokeratin 7 (cytokeratin 7), cytokeratin 20 (cytokeratin 20), melanoma-associated antigen (MART1), tyrosinase (tyrosinase), P-ALP (PLAP), liver cell specific antigen (hepatocyte specific antigen), flesh specificity actin (muscle specific actin), prostate specific acid phosphatase (PSAP), flesh forms albumen (myogenin), synaptic vesicle albumen (synaptophysin), differentiation group CD7, CD10, CD4, CD8, CD21, CD23, terminal deoxynucleotidyl transferase (terminal deoxynucleotidyl transferase TdT), calcium (looking) nethike embrane albumen (calretinin), the nephroblastoma (WT) and thyroid gland transcription factor (TTF-1).

28. according to the method in the claim 27, it is characterized in that, wherein said one group of detection target more comprises: cytokeratin 5, cytokeratin 6, cytokeratin 8, cytokeratin 14, cytokeratin 18, cytokeratin 19, inhibin, mammary gland globin (mamaglobin), P27 albumen, terminal deoxynucleotidyl transferase (terminal deoxynucleotidyl transferase) TdT, anti-tartaic acid phosphatase tartrate resistant acod phosphatase (TRACP 5b), alpha-fetoglobulin (AFP), human chorionic gonadotropin (hCG), epithelial growth factor receptor (EGFR), zeta chain GAP-associated protein GAP-70(Zap-70), P63 albumen, differentiation group CD1a, CD34, CD35, CD43, CD56, CD57, DC79a, B cell lymphoma albumen-6(Bcl-6), degeneration lymthoma kinases anaplastic lymphoma kinase-1 (Alk-1), actin (fascin), B clone idiosyncratic transcription factor Pax-5, P504s albumen, positive Cycloxygenase-2(Cox-2), adenovirus, cytomegalovirus (CMV) Ags, Epstein-Barr virus (EBV) Ags, pylorus bacteroides antigen (Helicobactor pylori Ags), human papilloma virus humam papillomavirus (HPV) Ags, hepatitis B NA (HBcAg), herpes virus hominis I type and II type (HSVI ﹠amp; II Ags), transcription factor (c-myc gene translocation), polyadenous purine tail messenger RNA (poly A mRNAs), immunoglobulin (Ig) κ messenger RNA (Ig kappa mRNA) and immunoglobulin (Ig) λ messenger RNA (Ig lambda mRNA) in breast cancer Her2 GFP amplification (Her2gene amplification), the nuclear.

29. the method according in the claim 25 is characterized in that, described cellular array further comprises and is derived from melanomatous cell line.

30. the method according in the claim 29 is characterized in that, described cellular array further comprises the cell line that is derived from sarcoma.

31. the method according in the claim 30 is characterized in that, described cellular array further comprises the cell line that is derived from pernicious hematopoietic tissue.

32. the method according in the claim 31 is characterized in that, described cellular array further comprises the cell line that is derived from nervous system neoplasm.

33. according to the method in the claim 25, it is characterized in that, be characterised in that and comprise the following step that is embedded between steps d and the step e: handle the cell of the in vitro culture of results with chemical reagent, used chemical reagent is select from formaldehyde, acetaldehyde, alcohol, acetone, acetic acid, dimethylbenzene, dimethylbenzene substitute and paraffin.

34. the method according in the claim 25 is characterized in that, it is immunohistochemistry, immunocytochemistry or in situ hybridization that described pathology detect.

35. the method according in the claim 25 is characterized in that, wherein said evaluating objects comprises protein, DNA sequence and RNA order.

36. according to the method in the claim 25, it is characterized in that, wherein said one group is detected in the target and comprises the detection target that comes from microorganism, and simultaneously, wherein at least one is included in the detection target that cell line in the described quality control cellular array is expressed described microorganism.

37. the method according in the claim 25 is characterized in that, further comprises with inert material paraffin or wax to cover and immerse the cell that invests substrate surface

38. a right to use requires the method for the device in 19, is characterised in that:

A. on described quality control apparatus, to described one group of target that detects in the target, use particular agent, carry out analytical procedure,

B. by observation, imaging or digital scan log,

C. for each is included in cell point in the described cellular array, negative positive according to signal, a signal signature is specified in signal intensity and subcellular location,

D. to each cell point in the described cellular array, described test signal sign and described database are compared.

39. pathological analysis quality control kit, it is characterized in that, comprise a quality control cellular array on the solid substrate on plane, one comprises one group of media and media that comprises the data bank that pre-establishes that detects the component identification in the target; This data bank that pre-establishes detects target with described one group and described quality control cellular array connects.