CN103347574B

CN103347574B - The cellular array quality control apparatus of pathology morphological analysis

Info

Publication number: CN103347574B
Application number: CN201080034906.5A
Authority: CN
Inventors: 朱伟星
Original assignee: Individual
Current assignee: Individual
Priority date: 2010-12-07
Filing date: 2010-12-07
Publication date: 2016-04-20
Anticipated expiration: 2030-12-07
Also published as: CN103347574A; WO2012078138A1

Abstract

The present invention announces a pathological analysis quality control apparatus based on cellular array and making and use method thereof.The cultured cell in vitro of kinds cancer type and origin is placed on solid substrate by this device, forms low-density cellular array.This device can be immunity and molecular assay program (such as immunohistochemistry, immunocytochemistry and in situ hybridization) provides display multi-functional Quality Control that is positive and negative reaction to indicate.

Description

The cellular array quality control apparatus of pathology morphological analysis

Technical field

The invention relates to and manufacture and use the method being loaded with the Quality controlled slides of low density cellular array.Because the characteristic of the culturing cell on slide glass is known maybe can being known 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 that cellular array quality control slide glass can replace the tissue being usually difficult to obtain to make controls slide.

Background technology

Pathology detect, and such as IHC, ICC, and ISH, be used on patient specimen and do routine cancer diagnosis and prediction.General IHC, ICC and ISH detect need into many steps, comprise acquisition biological sample, fixed preparation in formalin, process sample, to reach dehydration and transparent object, specimen embedding in paraffin, is serially cut into slices and is loaded on microscope slide.Next also to carry out dewax (at dimethylbenzene, processing in alcohol and water), finally carry out various reaction.Reaction comprises a series of solution, and as enzyme, first antibody, second antibody, detection reagent, developer, drips on tissue slice, and insulation is wash-out also.After having reacted, tissue slice is placed in basis of microscopic observation.

Traditional biopsy sample microscopic examination display be overall biological cells and tissues framework.Such as, under conventional bush and eosin stains (H & E), nucleus is dyed to purple (brazilwood dyeing), and tenuigenin is dyed to redness (eosin stains).But, be usually not enough to do diagnose accurately from the H & E information drawn that dyes.The result that IHC and ISH detects can expand to molecular level to the cell size of tissue samples and shape analysis.The expression of specific albumen and gene in tissue samples can be detected with antibody and nucleic acid probe.Whether the existence of some detection target or biomarker can show the pedigree of tumour to contribute to diagnosis or the prognosis to specific antitumor therapy.In addition, these methods also can be used for detecting microorganism, as bacterium, and fungi, and virus.

Most of Pathology Lab is with determining that the tissue sample containing specific antigen makes the positive control of IHC in advance.This needs the tissue sample to a group is used as IHC positive control to register, section and file.Need preparation new when assaypositive tissue sample is finished.In daily IHC testing process, each antibody needs a positive tissue control.So each antibody is all verified in a certain concentration.

Improve as one, Battifora (United States Patent (USP) 4,820,504and5,610,022) has set forth and multiple assaypositive tissue sample (mostly being tumor tissues sample) has been embedded in a paraffin mass together, while method in contrast.This " many tissue tumors wax stone " simplifies the slicing processes of positive control tissue.The people such as Furmanski have set forth one diverse ways preparation " many tissue tumors wax stone " (U.S.Pat.No.4,914,022) slightly.Their improvement is that the core of organizing cutting out from multiple fundamental weave with common beverages suction pipe is embedded in a paraffin mass.

But using-system causes very large burden in contrast histology technician, because excess tissue sample must be collected, preparation contrast slide, and their antigenic characteristic is analyzed.Many investigators used non-organization material, the cell even polypeptide of such as vitro culture.The manufacturer of many investigators and IHC diagnostic kit contrasts the cell of known dyeing property point on slide glass as positive or negative.The people such as Moskaluk (C.A.MoskalukandM.H.Stoler, DiagnosticMolecularPathology, 2002, p234-238) have set forth culturing cell to be embedded in agarose and have made polyclonal cellular array and be placed on slide glass.Zhu (United States Patent (USP) 7,598,036) has set forth protein, nucleic acid, or cultured cell in vitro is directly put on slide, or first to put on special film to be attached on microscope slide together with detection sample by film again and be used as external control.Set forth a kind of method contrasted as IHC in order to the polypeptide of covalent bond on solid support.

Summary of the invention

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

One aspect of the present invention is Quality control cells array.The region (point) of multiple independent dispersion that cellular array is made up of various kinds of cell is formed.The cell that cellular array comprises can be normal cell, also can be optimum or malignant cell.An embodiment of Quality control cells array comprises origin to come from kinds of tumors or cancer type (carcinoma cancer, melanoma melanoma, sarcoma sarcoma, leukemia/lymphoma leukemia/lymphoma, andtumorsoftheneuralsystem and nervous system neoplasm) the cell the separated point of culturing cell composition.The mixture of a kind of cell or several cell is by o'clock on the surface of a solid support independently region, and this region is called as cell point.Cellular array is made up of multiple cell point, can be placed in microscope slide surface or be placed in can be attached to micro-surface of glass slide film or film on.

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

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

Another embodiment of the invention is each clone in Quality Control cellular array is positive control to some detection, is negative controls to other detections simultaneously.Another embodiment of the invention is the cell in Quality Control cellular array is that vitro culture obtains, and through chemical reagent process.The chemical reagent of process cell comprises formaldehyde, valeral, ethanol, acetone, acetic acid, dimethylbenzene, or dimethylbenzene surrogate.Another embodiment of the invention is that Quality Control cellular array comprises and is less than 50 cell points.Another embodiment of the invention is containing 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 Quality Control cellular array is put on microscope slides.Described microscope slides has a marked region and one to comprise the surveyed area of Quality Control cellular array.Another embodiment of the invention is that Quality Control cellular array is put on microscope slides.Described microscope slides has a marked region, a sample areas, and another one comprises the region of Quality Control cellular array.Another embodiment is a label comprising patient or sample information being attached to mark zone, the predetermined expression general layout of a certain detection target on Quality Control cellular array be imprinted on label, and (selectable) detects the date.Another embodiment is that Quality Control cellular array device comprises further and determines that one group by the target detected by the detection method of Quality Control.Another embodiment is the data set that Quality Control cellular array device comprises that a predetermined contact detects target and Quality Control cellular array further.

Another aspect of the present invention is the method manufacturing Quality Control cellular array device.First, the target that one group of detection method of carrying out Quality Control by Quality Control cellular array device detects is determined.According to the detection target group selected, determine the clone be contained in cellular array.Detect each target in target group should at least at one by positive expression in the clone selected.Alternatively, all members detected in target group have at least a clone to be negative expression in selected clone.Detect target to refer to be qualitatively or quantitatively determined in the cell that detection sample and Quality Control cellular array comprise presence or absence protein, gene, RNA molecule.Detecting step refers to immunohistochemical methods, immunocytochemistry, and in situ hybridization.For immunohistochemical methods 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 detects target to be as follows included in target group: pencytokeratins, EMA, S-100, HMB45, smoothmusclespecificantigen, ER, PR, Her2, P53, PSA, vimentin, chromogranin, desmin, CD3, CD20, CD117/c-kit, CD15, CD30, CD45, CD99, Bcl-1 (cyclinD1), Bcl-2, Ki-67, Igkappa, Iglamda, and CEA.According to our research, above 26 cell proteins are biomarkers that daily immunohistochemical methods or cellular immunization are the most frequently used in testing clinically.Their composition Quality Control cellular arraies carry out the Spot detection target group of quality control.

Another aspect of the present invention is the method using Quality Control cellular array device.Comprise and set up one for the predetermined data set of cellular array and relevant detection target group; Data set is presented in paper media Jie or electronic media; On test sample and Quality Control cellular array, carry out test procedure with the reagent of the member in test detection target group simultaneously; To observe, imaging, or test result is recorded in digital scanning; And by the Data Comparison in test signal and predetermined database.

An embodiment of the invention are predetermined data sets is determined by following steps: on Quality Control device, carry out test procedure to each member detected in target group, to observe, imaging, or test result is recorded in digital scanning, and target is detected to each for each the cell point in Quality Control cellular array.Embodiment is the predetermined data set of Quality Control device is be stored in digital media as in CD or DVD.In this case, data can show or be printed on paper on computer monitor.Another embodiment is that predetermined data set can be rendered as form, diagram, or one group of image.

Accompanying drawing explanation

Fig. 1 illustrates the sample of a Quality control cells array apparatus.Quality control cells array is an indicia on microscope slides 1, and (Fig. 1 a) wherein to comprise a sample areas 2, control area 3 and a marked region.Fig. 1 b points out that a label printed is attached to the label area of microslide.This label 5 printed comprises sample ID6, concrete analysis sample ID7, one that Quality control cells array the is made a concrete analysis of target display sample 8 decided in advance, and any one analyzes the database 9 of display.Fig. 1 c illustrates the signal designation in the test result of Quality control cells array.

Fig. 2 illustrates 9 positions possible on the Quality control cells array of microslide.The edge close to microslide is positioned at a, b, c and d cellular array.The later test sample book on same slide glass of orientating as of these cellular arraies leaves enough spaces.The central authorities of microslide are positioned at these cellular arraies of e and f.Slide glass e and f does not leave sample areas.

Fig. 3 is the sample of a Quality control cells array.Ten cell points are aligned to these cellular arraies of two rows 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 be for Quality control cells array set up a predetermined database outline demonstration.On Quality control cells array, carry out IHC or ISH by antibody or probe for objectives to analyze, be included in detection target group and be controlled by quality cellular array control.This analytical results reads by naked eyes or machine the microslide carrying Quality control cells array and carrys out record, and is stored on paper or on digital media.Quantitative or semiquantitative instruction is all assigned in test signal in test result.These programs need to be repeated, and each evaluating objects is included in and detects in target group, are used for setting up a Quality control cells array and the database relevant with detecting target group.

Fig. 5 illustrates functional quality in IHC, ICC or ISH analyze and controls the step of cellular array analysis.IHC, ICC or ISH analyze and carry out in test sample book with on Quality control cells array in test sample book simultaneously, under same condition by antibody or probe for interested target, be included in detect in target group and be controlled by quality cellular array and control.This analytical results, that is, this signal pattern, produce observed on Quality control cells array and compared by with the information in the database decided in advance.Coupling represents the test success of this sample.Unmatch list shows the test crash of this sample.

Fig. 6 illustrates and sets up and use and Quality control cells array and the program detecting the relevant computer software of target group and database.Two parallel information streams are required.First information stream is that the ID of manual typing evaluating objects or antibody/probe enter computer software.Second information stream is facilitated by machine, needs the image of painted cellular array and inputs target or antibody/probe I D, passing through machine by readable digital marking (such as barcode).For these two information streams, computer software is that each test antibody/probe distributes an ID (such as numeral or barcode), relevant with the painted areas of specific antibodies/probe, in the database decided in advance of cellular array, be stored in computer, and by analyzing decimal system software.This computer is that specific objective or antibody/probe print painted pattern, for manual program, is namely used for contrasting with the painted pattern in Realistic Analysis by surgical staff.In machine stream, the rendered image of needs is contrasted by analysis and measurement and the image in the database decided in advance, then confirms that result will be generated.This slide glass readout device can be installed on automatic staining device and realize automated quality control.This slide glass readout device also can be used as an automated quality and control proofreading equipment.

Embodiment

The present invention relates to as immunohistochemistry, cytology and molecular pathology Epidemiological Analysis provide the device of quality control, and use this device to analyze the detected material in cell or tissue section, especially, during immunohistochemical test, the method for quality control is realized.It is important to note that the present invention relates to Fabrication quality control device, comprise the method 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 the region of multiple independent dispersion that is made up of dissimilar culturing cell, and these cells are fixed and adhere to the planar solid phase carrier surface through agent treated.Before this planar solid phase carrier can be divided into (also can in this, as above) and below (also can in this, as bottom surface).This planar solid phase carrier can be the detection platform (e.g., for fractographic slide glass) of a plane, also can be the film adhered to by the tackiness agent of bottom surface in detection platform.

1. Quality control cells array apparatus

An aspect of this invention is Quality control cells array, the region (point) of the multiple independent dispersion be namely made up of various kinds of cell.Mentioned herein to cell can be normal cell, also can be optimum or malignant cell.Can be the cell be directly separated from tissue or body fluid, also can be culturing cell.Staging (type) comprising: cancer, melanoma, sarcoma, lymphoma/leukemia and neural system knurl.The mixture of the culturing cell of single kind or two to three kinds of dissimilar culturing cells, is loaded the isolated area be separated from each other in surface of solid phase carriers, is referred to herein as cell point.

Professional term, cancer, sarcoma, leukemia/lymphoma, myelomatosis and melanoma, be commonly used to the tumour distinguishing different tissues origin.Cancer, is that one originates from epithelial tumor type, is modal cancer type, accounts for greatly the 80-90% of all Malignant Tumor Cases.Sarcoma is a kind of tumor type originating from muscle, bone, cartilage, fat or reticular tissue.Leukaemia origin is in white cell and precursor cell thereof.Lymphoma is the malignant tumour originating from bone marrow derived cell, and involves lymphsystem.Leukemia and lymphoma are all the malignant tumours of hemopoietic system.In the application's book, leukemia and lymphoma are collectively referred to as Hematopoietic Malignancies.Melanoma originates from melanocytic malignant tumour, and melanophore can be produced as the painted melanochrome of skin, hair, eyes.

One of concrete scheme of quality control apparatus (microslide 1 that Fig. 1 is a) special, it has by the quality control district being multiplely dispersed in distribution, separate cell point 3 is formed, each cell point forms by individual layer or close to the culturing cell of individual layer, and the mixture of often kind of cell or two to three kinds of cells is restricted in individual cells point.Generally, cell adheres to carrier surface by covalent attachment.When organization need is attached at the slide glass of electric charge process, cell also can adhere to carrier surface by charge effect.This slide glass also comprises a sample areas 2 and a label area 4.The concrete scheme two of quality control apparatus, the label (Fig. 1 b) of slide glass, it comprises a sample identification marker 6, target (or antibody/probe) identification marking 7, label is also printed on the expression pattern 8 of predetermined particular target in cellular array, this writes specific expression pattern also can show 9 by the routine analyzer of Quality Control cellular array, can also the region of selective marker date of test.The concrete scheme three of quality control apparatus, provides a series of and pre-determines, the diagram (Fig. 1 c) of the expression pattern in cellular array 8 of various target.This diagram comprises, but be not limited to following three: (A) is filled with the solid dot of different uneven color, for Fig. 1 cA, the lightest point represents background stainings (maybe target/analyte can not do not detected), along with the intensification of solid dot fill color, represent critical expression (+/-) step by step, the weak positive (+), positives (++), and strong positive (+++); (B) Subcellular Localization of detected target is used to indicate with the letter of circle, for 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 be positioned at nucleus, there is the alphabetical c of circle to be designated as positive signal and be positioned kytoplasm, have the m of circle letter to be designated as positive signal and be positioned cytolemma; (C) circle changed, for chart 1cC, the circle that dotted line surrounds is designated as background signal (feminine gender), in the circle that solid black lines surround, adularescent point is designated as tenuigenin signal, have black point to be designated as nuclear signal in white circle, cover has the circle of a circle heavy black line bar to be designated as cytolemma signal.

The concrete scheme four of quality control apparatus, surface adhesion has the film of multiple independent cell point, and the bottom surface of this film can adhere to the test surfaces of selected detection platform, such as, microslide.

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

The concrete scheme five of quality control apparatus, polytype cell is embedded in the wax stone making in paraffin and include various kinds of cell type.These cell blocks can be made into section further, and adhere to surface of solid phase carriers.A kind of method making various kinds of cell type wax stone is that cell that is fixing and that processed is made into the columniform core of a branch of bundle, and is embedded in paraffin by them; Another kind method is, in the kapillary that can be cut off by microtome, fix and process the cell of particular types, multiple such kapillary assembles bunchy, and is embedded in paraffin.Quality control cells array can be placed on same surface of solid phase carriers with sample or be positioned on independent carrier.This carrier can be can control and/or detect the microslide of sample, transparent film, film or cover glass by sticking quality.Transparent film or film can by various macromolecular materials, as nylon, polystyrene, polypropylene, cellulose nitrate etc. are made with glass.The solid phase plane of film or film preferably 0.001mm to 0.5mm thickness.

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

2. select the clone being used for Quality control cells array

One of concrete scheme, by determining the clone in Quality control cells array to the detection of some row targets.Preferred plan is, forms cellular array by the least possible cell point, can carry out quality control again to clinical correlation analysis as much as possible simultaneously.Here, analysis refers to and detects specific analyte, target or biomarker.Such as, detecting a kind of protein target with a kind of antibody by immunohistochemical method is an analysis, and detecting another kind of protein target with another kind of antibody by the method for immunohistochemical methods chemistry is exactly another one analysis.

A kind of clone refer to single origin, purified culturing 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 coming from human malignancies are included in ATCC catalogue.Derive from that the clone of tumor tissues or cancerous tissue is many to be obtained by Secondary Culture or virus transfection, therefore they can express the antigen that great majority are expressed in primary tumo(u)r or cancer.By standardized cultural method, be easy to the culturing cell obtaining a large amount of different clone.

Culturing cell comprises thousands of kinds of different proteantigens and other biomolecules.But, in thousands of kinds of different protein and biomolecules, only have minority to have value to clinical diagnosis and Index for diagnosis.These molecules are commonly called biomarker.In a lot of culturing cell, the phraseology of biomarker is similar to its expression in pathological tissues cell.The clone in the present invention with potential using value derives from Several Kinds of Malignancy, and therefore, expression or the disappearance of its clinical relevant biomarkers are similar to the expression of tumour cell in these malignant tumours.

Concrete scheme two, Quality control cells array can provide the target of control test to contain a series of biomarker with very high clinical reference value.Through qualification, 26 kinds of biomarkers are had to have very high clinical reference value, comprise Keratin sulfate (pancytokeratins), EMA, S-100, HMB45, unstriated muscle specific antigens (smoothmusclespecificantigen), estrogen receptor (estrogenreceptor, ER), progesterone receptor (progesteronereceptor, PR), Her2, P53, prostate specific antigen (prostatespecificantigen, PSA), vimentin (vimentin), chromogranin (chromogranin), desmin (desmin), CD3, CD20, CD117/c-kit, CD15, CD30, CD45, CD99, Bcl-1 (cyclinD1), Bcl-2, Ki-67, Igkappa, 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 constitute the core that Quality control cells array can provide the target of control test.

Concrete scheme three, Quality control cells array can provide the target easily extensible of control test, contain following cell protein: high molecular weight cell Keratin sulfate (cytokeratin, HMW), lower molecular weight cytokeratin (cytokeratin, LMW), CK7 (cytokeratin7), CK20 (cytokeratin20), EMA, S-100, HM45, MART-1, tyrosine oxidase (tyrosinase), PLAP, liver cell specific antigens (hepatocytespecificAg), flesh specificity muscle fibrin (musclespecificactin), unstriated muscle specific antigens (smoothmusclespecificantigen), ER, PR, Her2, P53, PSA, PSAP, Myogenin (myogenin), chromogranin (chromogranin), synaptic vesicle proteins (synaptophysin), desmin (desmin), vimentin (vimentin), CD99, CD3, CD7, CD10, CD4, CD8, CD20, CD117/c-kit, CD5, CD15, CD21, CD23, CD30, CD45, Bcl-1 (cyclinD1), Bcl-2, Ki-67, Igkappa, Iglamda, terminal deoxynucleotidyl transferase (terminaldeoxynucleotidyltransferase, TdT), calcium (depending on) nethike embrane albumen (calretinin), WT-1, TTF-1, and CEA.

Concrete scheme four, Quality control cells array can provide the target of control test to further expand, contain following cell protein: high molecular weight cell Keratin sulfate (cytokeratin, HMW), lower molecular weight cytokeratin (cytokeratin, LMW), CK5 (cytokeratin5), cytokeratin 6 (cytokeratin6), CK7 (cytokeratin7), CK8 (cytokeratin8), CK14 (cytokeratin14), CK18 (cytokeratin18), Cyfra21-1 (cytokeratin19), CK20 (cytokeratin20), EMA, statin (inhibin), mammaglobin (mamaglobin), Ki-67, P27, P53, terminal deoxynucleotidyl transferase (terminaldeoxynucleotidyltransferase, TdT), , the Phosphoric acid esterase of resistance to tartaic acid (tartrateresistantacidphosphatase, TRACP5b), TTF1, PLAP, AFP, hCG, estrogen receptor (estrogenreceptor, ER), Her2, EGFR, progesterone receptor (progesteronereceptor, PR), Zap-70, P63, prostate specific antigen (prostatespecificantigen, PSA), prostate specific acid phosphatase (prostatespecificacidphosphatase, PSAP), liver cell specific antigens (hepatocytespecificAg), chromogranin (chromogranin), synaptic vesicle proteins (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 lymphom kinase-1 (anaplasticlymphomakinase-1, Alk-1), fascin, IgKappa, IgLamda, Pax-5, P504s, MART-1, HMB45, S-100, PAP, NF, tyrosine oxidase (tyrosinase), calcium (depending on) nethike embrane egg (calretinin), WT-1, CEA, Cox-2, flesh specificity muscle fibrin (musclespecificactin), unstriated muscle muscle fibrin (smoothmuscleactin), desmin (desmin), Myogenin (myogenin), with vimentin (vimentin).

Concrete scheme five, Quality control cells array can provide the target of control test to further expand, contain following cell protein and biomarker: flesh specificity muscle fibrin (musclespecificactin), unstriated muscle muscle fibrin (smoothmuscleactin), 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, chromogranin (chromogranin), TRPM-2 (clusterin), high molecular weight cell Keratin sulfate (cytokeratin, HMW), lower molecular weight cytokeratin (cytokeratin, LMW), CK5 (cytokeratin5), cytokeratin 6 (cytokeratin6), CK7 (cytokeratin7), CK8 (cytokeratin8), CK14 (cytokeratin14), CK18 (cytokeratin18), Cyfra21-1 (cytokeratin19), CK20 (cytokeratin20), desmin (desmin), EMA, fasciclin (fascin), granzyme B (granzymeB), HBME, oxyphorase (hemoglobin), IgA, IgD, IgE, IgG, IgM, J-chain, Ki-67, Igkappa, Iglambda, LMP-1, MUM1, myeloperoxidase (myeloperoxidase), NSE, AbOCT-2, S-100, Pax5, TDT, TIA-1TRAP, TTF-1, Vs38c, vimentin (vimentin), Zap-70, CD83, P62, CK18, CD163, P63, P16, Stat-3, calcium (depending on) nethike embrane egg (calretinin), BerEp4, CK5/6, ubiquitin (ubiquitin), tyrosine oxidase (tyrosinase), PR, ER, synaptic vesicle proteins (synaptophysin), CAM 120/80 (E-cadherin), MyoD1, ommatidium associated transcription factor antibody (microphthalmia), MUC-1, MUC-2, ln (laminin), Myogenin (myogenin), HER2, EGFR, MOC-31, CD99, TAG-72, MLH-1, MSH-2, AR, villin (villin), melanic related antigen (melanomaassocAg), ACT, DN5 (alpha-1-antitrysin), P27, P504s, statin (inhibin), PSA, AFP, MUC-5, MUC-6, P21, fibronectin (fibronectin), P53, beta-catenin (beta-catenin), CD123, CD24, CD133, Adenovirus Antigen (adenovirusantigens), cytomegalovirus antigen (cytomegalovirus (CMV) antigens), Epstein-Barr virus antigen (Epstein-Barrvirus (EBV) antigens), Heliobacter pylori antigen (Helicobactorpyloriantigens), Human papilloma virus HPV antigen (humanpapillomaviruses (HPV) antigens), HBcAg, HSVI & IIantigens, Her2 gene amplification (Her2geneamplification), c-myc gene dystopy (c-mycgenetranslocation), poly-AmRNA, IgkappamRNA, and IglambdamRNA.

Concrete scheme six, detects target group and can only include one or more microbial antigens or gene.In the case, the cell in the cell point that comprises of Quality control cells array comprises antigen or the gene of one or more microorganisms.Meanwhile, Quality control cells array also can comprise the cell point that does not contain microbial antigen or gene, as negative control.

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

Except the albumen target involved by immunohistochemistry IHC, the nucleic acid target that Quality control cells array also can be in cross experiment ISH in position provides contrast.As, c-myc gene dystopy, Her2 gene amplification, poly-ARNA and kappa/lambdamRNA is exactly the contrast of In situ hybridization ISH.

In immunohistochemistry IHC, detecting target group is proteinaceous antigen, detects by specific antibody.In the present invention, detect target group and can comprise all above-mentioned marks, even more, e.g., the biomarker of the useful clinical value of new discovery and introducing.In this case, Quality control cells array must can comprise the culturing cell system of greater amt.According to specific diagnosis, Index for diagnosis or research needs, the detection target group of reducing targetedly can be worked out.At this moment, structure meets the cellular array clone that just only needs are relatively less of Quality Control requirement.

For realizing the quality control to selected specific detection target group, in group, each composition should at least also can be detected, to provide positive control expressed by a kind of clone in constructed cellular array; In group, each composition also should at least can not be detected, to provide negative control not expressed by a kind of clone in constructed cellular array.But if detect the detection target or the marker that include wide expression in target group, cellular array just can not provide negative control for this target.The detection target of wide expression or marker, as: PCNA, GAPDH, beta-actin and much other albumen (house-keeping gene associated protein) participating in the basic physiological function Metabolism regulation of cell can be expressed in all cells system.The concrete scheme addressed this problem is, adds the clone of a non-human in cellular array, as the negative control of human biological's marker of wide expression.In this invention, the concrete scheme of cell line selection also comprises, and in Quality control cells array, for the detection target of a specific non-wide expression, be no more than 75%, 50%, 25%, or the cell point of 10% can produce the positive signal that can be detected.This invention is also embodied in, and in detection target group, the target of wide expression is indicated, and is different from other targets in group.Noun " detectable level " refer to expression or the existence that the strength of signal produced in the detection can show a certain target or marker, can be color or fluorescent signal; " detectable level " refer to higher than background noise strength of signal." undetectable level " refer to the strength of signal that can not distinguish with background noise.

A concrete scheme of this invention is that the cell point that the clone adding a non-human in Quality control cells array is formed, comes, for all targets, to comprise the target of wide expression, provide negative control.Here, the clone of non-human is not limited to certain specific non-human's clone.The researchist of specialty thinks, according to the needs of particular detection target group, from the clone all used in the present invention cellular array of multiple organism as: mouse, ox and insect.

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

Detect target group and an origin can come from target (albumen or the gene order) composition of microorganism, also can both comprise microorganism target, and comprise again the target deriving from cell.The target deriving from microorganism can by introducing in cell point with microorganism direct infection culturing cell, also can by engineered method microbial gene or gene fragment be imported and express in cell.

3. the using method of Quality control cells array

This invention also comprises device using method as described herein.Cellular array Quality Control slide glass can be used alone, or the test zone of point on slide glass, and its test zone can load sample to be tested, and (Fig. 1 a).Confirm and verify the reliable method of test-results of immunohistochemistry IHC, immunocytochemistry ICC, in situ hybridization ISH, comprise the target detected in biological specimen and whether exist and cellular localization.Method comprises, when detecting some or multiple target molecules, and the cellular array quality control apparatus in synchronous processing biological sample and the present invention.Here noun " process " used refer to perform and detect target molecules and whether exist and all programs of Subcellular Localization.As, " process " can be perform the program of immunohistochemical test IHC, by detecting the existence of target protein with the antibody of this albumen specific combination.Under suitable conditions, antibody and target protein specific binding, the signal (as: chrominance signal that color reaction produces) that the antibody that detection is combined with target protein again carries, detects that these signals can indicate the existence of target protein, can not detect that signal instruction target protein is not expressed.The researchist of specialty is very familiar with more such testing sequence.Citing part, we provide standard immunohistochemistry IHC and in situ hybridization ISH operating process.

Make this device can by correct use, be also required to be cellular array and relevant detection target and set up a vertical default database.The database preset will store with the form of paper material or digital media and provide.For setting up presetting database, Quality Control cellular array device needing to carry out lot of experiments, with related reagent, each component detected in target group independently being tested.Such as, detect immunohistochemical test's database of target groups for setting up 26 molecule cores, will on independently cellular array device, by specific antibody, according to the operating process provided in example 2, independent experiment is carried out to wherein each component.All cells in 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 in other cell point, all cells will produce negative signal.

Noun " positive signal " refer to the signal that the detection reagent for interested particular detection target produces, higher than background signal, when visual inspection or machine detect, be easy to distinguish with nonspecific background signal.Noun " negative signal " refer to the signal that the detection reagent for interested particular detection target produces, when visual inspection or machine detect, can not distinguish with nonspecific background signal.These signals will be recorded and analyze, and provide instruction that is digital or numeral.Such as, Strong positive signals can change high pixel into, or is designated as multiple "+" symbol.Such as, " +++ " represents epistasis, and " ++ " represents positives, and "+" represents the weak positive, and " +/-" represents the uncertain positive, and "-" represents negative.Except the strength of signal of the target expression level in reacting cells, in analytic process, can also show and record the Subcellular Localization detecting each component in target group.After obtaining the information detecting quantitative or semi-quantitative results and the Subcellular Localization expressed in all cells point of target group all components at Quality control cells array, we just establish required presetting database.Database stores by paper material, comprises but is not only: catalogue, chart or pictures.Database also can be stored by a digital media and be presented, such as computer monitor and CD/DVD.

As to detect in target group the target that comprises in the cell point of Quality control cells array, form typical positive signal, can be used as individual authentication, prove that the operation detecting target in biological sample is accurate.When biological sample produces negative findings, as non-coloring or fluorescent signal, such contrast seems particularly important.The positive findings that Quality control cells array produces can confirm that the above results is real negative findings, and the problem of nonprogrammed mistake or reagent quality causes.Because biological specimen and Quality control cells array are adhered on same microslide, and under identical condition, contact with same reagent, therefore form checking reliably.

Another concrete scheme comprises, and determines the analytical procedure of target molecule Subcellular Localization in biological specimen.Method comprises, and in immunohistochemistry IHC or In situ hybridization ISH, as mentioned above, processes biological specimen and Quality control cells array simultaneously.Result shows as target molecule specific subcellular area presence or absence in the cell of biological specimen and the cell of Quality control cells array.In the Subcellular Localization of the detectable signal that biological specimen target produces and its cell point in Quality control cells array produce signal Subcellular Localization compare.

Subcellular location includes, but are not limited to: nucleus, tenuigenin, cytolemma and nuclear membrane.Expression level and their Subcellular Localization of particular target are constant in the specific cells system of Quality control cells array, are namely determined and record when designing and making Quality control cells array.A certain testing sequence is carried out to biological sample and Quality control cells array, detect certain target molecules result can with pre-determine and record, in Quality control cells array, the Subcellular Localization of this molecule in cell and expression level compare.As tested result in Quality control cells array specifically and pre-determining and the data consistent of certain particular target record, prove that the operation tested specifically is successful, test the credible result in biological specimen specifically, can report be sent.As tested result in Quality control cells array specifically and pre-determining and the data of certain particular target record are inconsistent, then illustrate that the operation tested specifically has problems, the result of this test in biological specimen is insincere, can not send report.

The present invention also comprises a commercial test kit.Test kit is by Quality control cells array apparatus, can realize by array the information that one group of quality control detects target, pre-determine and record to comprise in Quality control cells array each the detection expression level of target and database of Subcellular Localization information in each cell point.The expression level of target can be come quantitatively by digital pixel or the quantity of "+" symbol carrys out sxemiquantitative.Such as, " +++ " represents strong positive, and " ++ " represents the positive, and "+" represents the weak positive, and " +/-" represents uncertain, and "-" represents feminine gender.Wherein also include one and have recorded each detection target at the form of the expression level of each point or computer file, and being obtained by scanning of printing or each detection target of digital version in the image result of the expression level of each point.

Example 1 is included in the selection of the clone in Quality control cells array and controlled detection target group has close relation.In this example, 11 clones are selected the Quality control cells array being made into 11 points, comprise Cell line Hela (adenocarcinoma of the uterine cervix), LS174T (colorectal adenocarcinoma), SW480 (gland cancer, colorectal), BT474 (breast duct cancer), MCF-7 (mammary cancer), 22Rv1 (prostate cancer), Ragi (B cell, Burkitt lymphoma), HuT78 (t cell lymphoma), Jurkat (T cell leukemia), SK-UT-1 (sarcoma), SW1353 (sarcoma), with CaC1 (melanoma).These 11 kinds of clones selected from Quality control cells array, contain the core group of 26 members of target protein, to a certain extent, each target protein is in this core group, in at least one clone, all indicate a detectable level, target protein shows not detectable level at least one clone simultaneously.

Example 2 immunohistochemistry (IHC)

In this example, biological specimen (comprising Quality control cells array) is by antibody treatment (primary with secondary), and processed gives chromogen color development, then finally gives counterstaining.

A. deparaffinization

When the tissue slice using paraffin to implant, deparaffinization must be performed, and be representatively through 52 minutes of taking turns dimethylbenzene and soak, twice 100% alcohol, the alcohol of one time 95% makes rehydration, then at room temperature air dried five minutes.

B. antigen obtains

Well-known in the skill of the specific antibody of the painted middle use of IHC, be need to obtain program pre-treatment formalin set tissue slice by an antigen, normally obtain in solution by heating or proteolytic ferment treatment at antigen.After antigen obtains, sample, comprises Quality control cells array, is hatched 20-30 minute at room temperature by with the normal sheep of mixing or horse serum that contain 5%.

C. antigen-reactive that is initial and secondary

Followed by the hatching of serum, sample is rinsed by with PBS.Prediluted initial antigen serum or antibody (about 150-200mu.L) are applied to each sample.Sample is at room temperature hatched 2-4 hour by by antigen serum or antibody, or hatching 2 hours in the moist environment of 40 degrees Celsius, or can hatch a whole night in the environment of the humidity of a room temperature.The sample on slide glass after hatching, will rinse 3 times by PBS in staining dish.

The secondary antibody (about 150-200mu.L) watered down is used in each sample on slide glass.This is hatched 30 minutes by the environment of the humidity of 40 degrees Celsius.Sample after hatching, will rinse 3 times by PBS in staining dish.

D. in, raw peroxidase activity removes

Sample on slide glass is then placed into one and contains in the 500mlPBS solution of 3% hydrogen peroxide and 1% sodiumazide, and at room temperature hatching removes interior raw peroxidase activity for 15 minutes.The sample on slide glass after being hatched by hydrogen peroxide PBS, will rinse 3 times by PBS in staining dish.

E. color development

The active set of ABC synthetics (VectorLaboratoriesInc., Burlingame, Calif.) by using PBS to dilute them.This active set (about 150-200mu.L) is applicable to each sample on slide glass.The hatching of ABC solution is in the environment of the humidity of 40 degrees Celsius 30 minutes.The sample on slide glass after hatching, will rinse 3 times by PBS in staining dish.

DAB solution is that the solution containing 30%H2O2 by adding 100mgDAB to 100mLPBS and interpolation 50.mu.L prepares.The DAB solution of about 150-200.mu.L is added in each sample on slide glass.Color development can be monitored by microscopic examination.Coloured precipitation will be formed on positive cell.Color started to manifest after 2-5 minute, usually in 10 minutes, reached enough imagings, but the hatching of 20-30 minute may need to be used to weaker colo(u)r atlas.In order to stop development, the sample on slide glass will rinse 3 times by deionized water in staining dish.

F. painted on the contrary

Sample on slide glass immerses in the phenodin of Harris 10-50 second, and cleans for three times by being dipped into deionized water, to be then immersed in 0.2% solution of ammonium hydroxide 30 seconds, and to clean for three times by being dipped into deionized water.They to immerse in 95% ethanol twice, each 2 minutes, next to immerse in 100% ethanol twice, each 2 minutes, and finally, this slide glass, by immersing in dimethylbenzene twice, cleans for each 2 minutes.

Example 3 in situ hybridization

In this example, biological specimen homogenous quantities controls cellular array (being also used as sample from now on to quote) on slide glass, 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 glass by putting into dimethylbenzene four times, each 5 minutes, in 100% ethanol twice, each 1 minute, and in 95% ethanol twice, within each 1 minute, carry out deparaffinization.The tissue slice slide glass of deparaffinization is then by and flushing cleaned containing RNaseBlock (BioGenex, SanRamon, Calif.) deionized water.

B. the Proteinase K process of mounted tissue samples

The fresh Proteinase K Solution watered down of about 150-200.mu.L is placed on each on the sample of slide glass, and at room temperature hatches 15 minutes.After digestion, the sample on slide glass is rinsed 5 minutes by RNaseBlock by the PBS of 500mL on staining dish.Sample dewaters by immersing in a staining dish following solution in succession: 500mL distilled water additional RNaseBlock10 second, the ethanol of 500mL50% adds RNaseBlock10 second, 500mL 95% ethanol 10 second, and 500mL100% ethanol 10 second.Whole slide glass at room temperature dry 5 minutes.

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

The hybridization solution comprising VITAMIN or digoxigenin labeled oligonucleotide probe is placed on the sample of each slide glass.This needs the solution of about 50-100.mu.L.Slide glass is placed in a baking box or on a heat block, with 95 degrees Celsius of 8-10 minute, changes the habit of nucleic acid.This step eliminates the hairpin loop of mRNA order or turn over glue.After the step changed one's skin, slide glass, by under 45 degrees Celsius, hatches a whole night in a wet environment.Then hybridization step, slide glass is cleaned 2 times by staining dish.SSC (standardized citrate solution) is lower 5 minutes at 37 degrees Celsius, next within lower 5 minutes, is rinsed at 37 degrees Celsius by the SSC at 1 times.Next this rinsed 30 minutes at 60 c by the SSC at 0.2 times.Finally, this slide glass is rinsed 2 times by PBS, each 2-5 minute.

D. signal detection

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

E. the application of secondary antibodies

The secondary antibody (about 150-200.mu.L) watered down in advance is used in each in the sample of slide glass, and hatches 30 minutes in the environment of the humidity of 40 degrees Celsius.After hatching, the sample on slide glass is cleaned 3 times on painted dish by PBS.

F. in, raw peroxidase activity removes

The PBS that sample on slide glass is placed into containing 500mL contains in the painted dish of 3% hydrogen peroxide and 0.1% sodiumazide, and at room temperature hatches 15 minutes.The sample on slide glass after being hatched by hydrogen peroxide PBS, is rinsed 3 times on painted dish by PBS.

The application of G.ABC synthetics " ELITE "

The active set of ABC synthetics by using PBS to dilute them.This active set (about 150-200mu.L) is applicable to each sample on slide glass, and hatches 30 minutes in the environment of the humidity of 40 degrees Celsius.The sample on slide glass after hatching, will rinse 3 times by PBS in staining dish.

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

DAB solution by add in 100mgDAB to 100mLPBS and add 50.mu.L containing 30% H2O2 prepare.The DAB solution of about 150-200.mu.L is added in each sample on slide glass.The sample that color development can be observed on slide glass by DAB under the microscope monitors.Coloured precipitation will be formed on positive cell.Color started to manifest after 2-5 minute, usually in 10 minutes, reached enough imagings, but the hatching of 20-30 minute may need to be used to weaker colo(u)r atlas.In order to stop development, the sample on slide glass will rinse 3 times by deionized water in staining dish.

I. counterstain

Sample on slide glass immerses in the phenodin of Harris 10-50 second, and cleans for three times by being dipped into deionized water.Then slide glass to be immersed in 0.2% solution of ammonium hydroxide 30 seconds, and cleans for three times by being dipped into deionized water.They to immerse in 95% ethanol twice, each 2 minutes, next to immerse in 100% ethanol twice, each 2 minutes, and finally, this slide glass, by immersing in dimethylbenzene twice, cleans for each 2 minutes.

Claims

1. a quality control apparatus for pathology detection, it is characterized in that, comprise cellular array, described cellular array is characterised in that and comprises multiple different cell; Wherein said cell attachment is on the surface of a solid substrate; Simultaneously, wherein said various kinds of cell is arranged to the cell point of many separation, and described cellular array comprises the cell strain being derived from cancerous tissue, the cell strain being derived from sarcoma, is derived from the cell strain of nervous system neoplasm, is derived from the cell strain of malignant hematogenous tissue.

2. according to the device in claim 1, it is characterized in that, described cellular array comprises the culturing cell containing microbial antigen or gene.

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

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

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

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

7. according to the device in claim 1, it is characterized in that, wherein said cell point comprises a pure specific culturing cell or one by the specific cell mixing of different sorts.

8. according to the device in claim 1, it is characterized in that, comprise the cell crossed by infected by microbes.

9. according to the device in claim 1, it is characterized in that, comprise demarcation one group further and detect target.

10. according to the device in claim 9, it is characterized in that, described one group of detection target is characterised in that and comprises following member: Keratin sulfate (pencytokeratins), EMA (EMA), acid calcium associated proteins (S-100), melanoma-associated antigen (HMB45), unstriated muscle specific antigens (smoothmusclespecificactin), estrogen receptor (ER), progesterone receptor (PR), Her2, P53, prostate specific antigen (PSA), vimentin (vimentin), chromogranin (chromogranin), desmin (desmin), differentiation group CD3 (Clusterofdifferentiation3), CD20, CD117/c-kit, CD15, CD30, CD45, CD99, B cell lymphoma albumen-1Bcl-1 (cyclinD1), B cell lymphoma albumen-2Bcl-2, nuclear antigen Ki-67, Mitochondrion IgG (Immunoglobulin) kappa, Mitochondrion IgG (Immunoglobulin) lamda, with carcinomebryonic antigen (CEA).

11. according to the device in claim 10, it is characterized in that, wherein said one group of detection target comprises following compositions: CK7, CK20, melanoma-associated antigen (MART1), tyrosine oxidase, P-ALP (PLAP), liver cell specific antigen, muscle special efficacy Actin muscle, 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 Transcription Factor (TTF-1).

12., according to the device in claim 11, is characterized in that, wherein said one group of detection target is more comprise following compositions: CK5, cytokeratin 6, CK8, CK14, CK18, Cyfra21-1, statin, mammaglobin (mamaglobin), P27 albumen, terminal deoxynucleotidyl transferase (terminaldeoxynucleotidyltransferase) TdT, Tartrate resistant acid phosphatase tartrateresistantacodphosphatase (TRACP5b), alpha-fetoglobulin (AFP), human chorionic gonadotropin (hCG), epithelial growth factor receptor (EGFR), zeta chain associated protein-70 (Zap-70), P63 albumen, differentiation group CD1a, CD34, CD35, CD43, CD56, CD57, DC79a, B cell lymphoma albumen-6 (Bcl-6), anaplastic lymphom kinases anaplasticlymphomakinase-1 (Alk-1), Actin muscle (fascin), B cell system idiosyncratic transcription factor Pax-5, P504s albumen, positive Transitional cell carcinomas (Cox-2), adenovirus, cytomegalovirus (CMV), Epstein-Barr virus (EBV), Helicobacter pylori antigen (HelicobactorpyloriAgs), human papillomavirus humampapillomavirus (HPV) Ags, hepatitis B nuclear antigen (HBcAg), herpes virus hominis I type and II type (HSVI & IIAgs), mammary cancer Her2 protein gene amplification (Her2geneamplification), transcription factor (c-mycgenetranslocation) in core, polyadenous purine tail messenger RNA(mRNA) (polyAmRNAs), immunoglobulin (Ig) κ messenger RNA(mRNA) (IgkappamRNA), with immunoglobulin (Ig) λ messenger RNA(mRNA) (IglambdamRNA).

13., according to the device in claim 9, is characterized in that, described one group of detection target comprises microbial antigen or gene.

14., according to the device in claim 13, is characterized in that, described microbial antigen or gene come from bacterium, fungi, or virus.

15., according to the device in claim 9, is characterized in that, comprise a database further, described database is determined according to the following steps:

I. a detecting step is carried out on such devices with the test reaction thing of a member in detection one group detection target,

Ii. by observation, imaging or digital scanning log,

Iii. corresponding to described one group of member detected in target, is the cell point in cellular array described in each, specifies a detection signal to indicate according to test-results,

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

16., according to the device in claim 15, is characterized in that, described detecting step comprises Immunohistochemical detection, Immuncytochemical detection and in situ hybridization and detects; Wherein said detection signal comprises negative positive signal, strength signal, and subcellular localization signal; Wherein said test signal indicates and comprises letter designation and shape sign.

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

18., according to the device in claim 17, is characterized in that, comprise a printed label further, for being attached to described mark zone, described printed label characteristics is the signal mode comprising a predetermined described cellular array.

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

20., according to the device in claim 19, 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, meanwhile, below described solid substrate, post tackiness agent, so that attached work is on other substrate.

The method of the device in 21. 1 manufacturing claims 1, is characterised in that:

A. one group of detection target needing quality control in a pathology detection is selected,

B. select all cells strain that need be included in described cellular array, wherein detect target described in each and exist at least one cell strain selected, and can the positive be detected,

C. described cell strain is cultivated in vitro,

D. cultured cells strain is in vitro gathered in the crops,

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

22. according to the method in claim 21, it is characterized in that, wherein said one group is detected that target comprises: Keratin sulfate (pencytokeratins), EMA (EMA), acid calcium associated proteins (S-100), melanoma-associated antigen (HMB45), unstriated muscle specific antigens (smoothmusclespecificactin), estrogen receptor (ER), progesterone receptor (PR), Her2, P53, prostate specific antigen (PSA), vimentin (vimentin), chromogranin (chromogranin), desmin (desmin), differentiation group CD3, CD20, CD117/c-kit, CD15, CD30, CD45, CD99, B cell lymphoma albumen-1Bcl-1 (cyclinD1), B cell lymphoma albumen-2Bcl-2, nuclear antigen Ki-67, Mitochondrion IgG (Immunoglobulin) kappa, Mitochondrion IgG (Immunoglobulin) lamda, with carcinomebryonic antigen (CEA).

23. according to the method in claim 22, it is characterized in that, wherein said one group of detection target more comprises: CK7 (cytokeratin7), CK20 (cytokeratin20), melanoma-associated antigen (MART1), tyrosine oxidase (tyrosinase), P-ALP (PLAP), liver cell specific antigens (hepatocytespecificantigen), flesh specificity muscle fibrin (musclespecificactin), prostate specific acid phosphatase (PSAP), Myogenin (myogenin), synaptic vesicle proteins (synaptophysin), differentiation group CD7, CD10, CD4, CD8, CD21, CD23, terminal deoxynucleotidyl transferase (terminaldeoxynucleotidyltransferaseTdT), calcium (depending on) nethike embrane albumen (calretinin), the nephroblastoma (WT) and Thyroid Transcription Factor (TTF-1).

24., according to the method in claim 23, is characterized in that, wherein said one group of detection target more comprises: CK5, cytokeratin 6, CK8, CK14, CK18, Cyfra21-1, statin, mammaglobin (mamaglobin), P27 albumen, terminal deoxynucleotidyl transferase (terminaldeoxynucleotidyltransferase) TdT, Tartrate resistant acid phosphatase tartrateresistantacodphosphatase (TRACP5b), alpha-fetoglobulin (AFP), human chorionic gonadotropin (hCG), epithelial growth factor receptor (EGFR), zeta chain associated protein-70 (Zap-70), P63 albumen, differentiation group CD1a, CD34, CD35, CD43, CD56, CD57, DC79a, B cell lymphoma albumen-6 (Bcl-6), anaplastic lymphom kinases anaplasticlymphomakinase-1 (Alk-1), Actin muscle (fascin), B cell system idiosyncratic transcription factor Pax-5, P504s albumen, positive Transitional cell carcinomas (Cox-2), adenovirus, cytomegalovirus (CMV) Ags, Epstein-Barr virus (EBV) Ags, Helicobacter pylori antigen (HelicobactorpyloriAgs), human papillomavirus humampapillomavirus (HPV) Ags, hepatitis B nuclear antigen (HBcAg), herpes virus hominis I type and II type (HSVI & IIAgs), mammary cancer Her2 protein gene amplification (Her2geneamplification), transcription factor (c-mycgenetranslocation) in core, polyadenous purine tail messenger RNA(mRNA) (polyAmRNAs), immunoglobulin (Ig) κ messenger RNA(mRNA) (IgkappamRNA), with immunoglobulin (Ig) λ messenger RNA(mRNA) (IglambdamRNA).

25., according to the method in claim 21, is characterized in that, described cellular array comprises further and is derived from melanomatous cell strain.

26., according to the method in claim 25, is characterized in that, described cellular array comprises the cell strain being derived from sarcoma further.

27., according to the method in claim 26, is characterized in that, described cellular array comprises the cell strain being derived from malignant hematogenous tissue further.

28., according to the method in claim 27, is characterized in that, described cellular array comprises the cell strain being derived from nervous system neoplasm further.

29. according to the method in claim 21, it is characterized in that, be characterised in that and comprise the following step be embedded between steps d and step e: with the cell of the vitro culture of chemical reagent process results, chemical reagent used is select from formaldehyde, acetaldehyde, alcohol, acetone, acetic acid, dimethylbenzene, dimethylbenzene surrogate and paraffin.

30., according to the method in claim 21, is characterized in that, it is immunohistochemistry, immunocytochemistry or in situ hybridization that described pathology detect.

31., according to the method in claim 21, is characterized in that, wherein said evaluating objects comprises protein, DNA sequence and RNA order.

32. according to the method in claim 21, it is characterized in that, wherein said one group of detection target comprises the detection target coming from microorganism, and meanwhile, wherein at least one cell strain be included in described Quality control cells array expresses the detection target of described microorganism.

33., according to the method in claim 21, is characterized in that, comprise further and to cover with inert material paraffin or wax and to immerse the cell investing substrate surface.

The method of the device in 34. 1 use claims 15, is characterised in that:

A., on described quality control apparatus, to described one group of target detected in target, by particular agent, analytical procedure is carried out,

B. by observation, imaging or digital scanning log,

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

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

35. 1 pathological analysis quality control reagents boxes, it is characterized in that, comprise a cellular array as claimed in claim 1 on the solid substrate of plane, one comprises the medium that a group is detected the component identification in target, and a medium comprising the database pre-established; This database pre-established detects target by described one group and described Quality control cells array connects.