CN100360925C - Slide detection method employing one-by-one comparison - Google Patents
Slide detection method employing one-by-one comparison Download PDFInfo
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- CN100360925C CN100360925C CNB2005100503073A CN200510050307A CN100360925C CN 100360925 C CN100360925 C CN 100360925C CN B2005100503073 A CNB2005100503073 A CN B2005100503073A CN 200510050307 A CN200510050307 A CN 200510050307A CN 100360925 C CN100360925 C CN 100360925C
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Abstract
The present invention relates to a slide detecting method in a one-by-one control mode, which belongs to the technical field of medical detecting methods. The present invention is characterized in that the method comprises the following steps: control substances corresponding to a detected object are selected for cell extracting processing; a cell control storing body is prepared; after the cell extracting processing of the control substances is finished, the cell control storing body is made in a storing medium, and the proportion of cell extract and the storing medium is from 1: 1 to 5; after slide detection is carried out, the cell control storing body is a little painted, coated or molten on each corresponding slide to simultaneously carry out detection, analysis or judgment with the detected object. The slide detecting method in a one-by-one control mode has the advantages of simple and convenient operation, and can guarantee the detecting conditions of the slides to be consistent. The accuracy of a detected result is effectively increased, necessity data are provided for relevant research work, accurate judging basis is provided for the selectivity of treating schemes, and the performance of unknown substances can be detected by control observation.
Description
Technical field
The invention belongs to the medical detecting method technical field, be specially and contrast the formula slide detection method one by one.
Background technology
In scientific research or clinical practice, histology, cytology, histocyte level, protein level, rna level, dna level, SABC, various in situ hybridization or original position PCR, so long as the detection of in-situ method all needs to set up positive control and blank by quality requirements.At present, except that minority reagent can be sought in sheet the own control, normally one group of detection lug can only be set up a positive control sheet in contrast by dicing method, but no matter be the testing conditions unanimity that machine operation or manual operations all can not guarantee every slide, all there are differences as the time of each step, the consumption of reagent, the degree of tiring or other human factor etc., the capital causes the accuracy of partial results to be difficult to judge, so its standardization issue is subjected to everybody attention day by day.
Summary of the invention
The object of the invention is to overcome the above-mentioned problems in the prior art, and design provides a kind of technical scheme that contrasts the formula slide detection method one by one, can effectively improve the accuracy of judged result.
The described formula slide detection method that contrasts one by one is characterized in that may further comprise the steps:
1) selects to contrast material accordingly, carry out cell extraction and handle with detected object;
2) body is preserved in the contrast of preparation cell: after the contrast material carries out the cell extraction processing, make the cell contrast and preserve body in preserving medium, cell extract is 1 with the ratio of preservation medium: 1-5;
Described preservation medium is whiteruss or solid paraffin, also can be homemade preservation liquid, preserve the preparation of liquid: get 0.05-0.1mol/L, PH and be 7.4 trishydroxymethylaminomethane 100ml, be warmed to 60-70 ℃, add high-quality gelatin 50-100mg, after the stirring and dissolving, be cooled to room temperature, add the 0.1-0.8g bovine serum albumin(BSA), add NaN
3200mg dissolving back is filtered and is made;
3) when carrying out the slide detection, above-mentioned cell contrast is preserved a little point of body, is coated with or melted on corresponding each slide, detect, analyze or judge with detected object.
The described formula slide detection method that contrasts one by one is characterized in that described contrast material is cultured cell, liquid sample or tissue specimen.
The described formula slide detection method that contrasts one by one, the cell extraction that it is characterized in that cultured cell is treated to directly and with pancreatin cell is digested from culture flask.
The described formula slide detection method that contrasts one by one, the cell extraction that it is characterized in that liquid sample is treated to liquid sample and adds that anti-coagulants carries out direct centrifuging, cell is obtained in washing, described cleansing solution is that 0.01mol/L, PH are 7.4 phosphate buffer or TRIS buffer, and anti-coagulants is heparin or citric acid.
The described formula slide detection method that contrasts one by one is characterized in that the cell extraction of tissue specimen is treated to tissue specimen flush away bloodstain, grinds, and the 100-400 eye mesh screen filters, and carries out cell punching processing, centrifuging, washing.
The described formula slide detection method that contrasts one by one is characterized in that carrying out after stale tissue specimen grinds in time fixing, and immobile liquid is 10-20% neutral formalin or 90-95% ethanol or cold acetone.
The described formula slide detection method that contrasts one by one, it is characterized in that tissue specimen grinds the back and adds the erythrocyte cracked liquid processing, the preparation of erythrocyte cracked liquid: take by weighing 3-4g ammonium chloride, 1-2g trishydroxymethylaminomethane, be dissolved in water and be diluted to 500ml, 0.22 the degerming of μ m membrane filtration, 4-6 ℃ of preservation.
The above-mentioned formula slide detection method that contrasts one by one, simple, convenient, the cell contrast is preserved a little point of body, is coated with or melted on corresponding each slide, detect, analyze or judge with detected object, guarantee the testing conditions unanimity of every slide, effectively improved the accuracy of testing result, for relevant research work provides data necessary, more the selection of therapeutic scheme provides basis for estimation more accurately, and can also detect the character of unknown materials by paired observation.
Embodiment
Contrast the formula slide detection method one by one, it is characterized in that may further comprise the steps: 1) select to contrast material accordingly with detected object, carry out cell extraction and handle, described cell comprises zooblast and microorganism, and described contrast material can be cultured cell, liquid sample or tissue specimen.The cell extraction of cultured cell is treated to directly and with pancreatin cell is digested from culture flask.The cell extraction of liquid sample is treated to liquid sample adding anti-coagulants and carries out direct centrifuging, washs and obtain cell, described cleansing solution is that 0.01mol/L, PH are 7.4 phosphate buffer or TRIS buffer, and anti-coagulants is heparin or citric acid.The cell extraction of tissue specimen is treated to tissue specimen flush away bloodstain, grinds, and 300 eye mesh screens filter, and carries out cell punching processing, centrifuging, washing.Carry out after stale tissue specimen grinds in time fixing, immobile liquid is 15% neutral formalin or 95% ethanol or cold acetone.It is more that tissue specimen contains red blood cell, grinding the back adds erythrocyte cracked liquid and handles the preparation of erythrocyte cracked liquid: take by weighing 3.735g ammonium chloride, 1.3g trishydroxymethylaminomethane, be dissolved in water and be diluted to 500ml, 0.22 the degerming of μ m membrane filtration, 4 ℃ of preservations.
2) body is preserved in the contrast of preparation cell: after the contrast material carries out the cell extraction processing, in preserving medium, make the cell contrast and preserve body, it is suspension that body is preserved in described cell contrast, it also can be solid, described preservation medium is medical whiteruss or solid paraffin, also can be homemade preservation liquid.Preserve the preparation of liquid: get 0.06mol/L, PH and be 7.4 trishydroxymethylaminomethane 100ml, be warmed to 65 ℃, add high-quality gelatin 90mg, after the stirring and dissolving, be cooled to room temperature, add the 0.7g bovine serum albumin(BSA), add NaN
3200mg dissolving back is filtered and is made.Cell extract and the proportioning of preservation liquid by 1: 3 are made cell suspension; Cell extract dewaters, carry out waxdip with whiteruss, solid paraffin by proportioning after the transparent processing handles, and its ratio was respectively 1: 3,1: 1, makes cell suspension respectively, solid is preserved in the cell contrast.
3) when carrying out the slide detection, above-mentioned cell contrast is preserved a little point of body, is applied on corresponding each slide, detect, analyze or judge with detected object.
Related centrifuging, carrying out washing treatment among the application, dehydration, transparent, waxdip are handled, and the preparation that cell suspension, cell contrast are preserved solid is general known technology, do not repeat them here.
According to this detection method, can develop contrast material at different testing goals, different detected objects, produce the cell contrast that can preserve for a long time and preserve body.When carrying out the slide detection, the cell contrast is preserved a little point of body, is coated with or melted on corresponding each slide, detect, analyze or judge with detected object, guarantee the testing conditions unanimity of every slide, effectively improved the accuracy of testing result, for relevant research work provides data necessary, more the selection of therapeutic scheme provides basis for estimation more accurately, and can also detect the character of unknown materials by paired observation.
Claims (7)
1. contrast the formula slide detection method one by one, it is characterized in that may further comprise the steps:
1) selects to contrast material accordingly, carry out cell extraction and handle with detected object;
2) body is preserved in the contrast of preparation cell: after the contrast material carries out the cell extraction processing, make the cell contrast and preserve body in preserving medium, cell extract is 1 with the ratio of preservation medium: 1-5;
Described preservation medium is homemade preservation liquid, preserve the preparation of liquid: get 0.05-0.1mol/L, PH and be 7.4 trishydroxymethylaminomethane 100ml, be warmed to 60-70 ℃, add high-quality gelatin 50-100mg, after the stirring and dissolving, be cooled to room temperature, add the 0.1-0.8g bovine serum albumin(BSA), add NaN
3200mg dissolving back is filtered and is made;
3) when carrying out the slide detection, above-mentioned cell contrast is preserved a little point of body, is applied on corresponding each slide, detect, analyze or judge with detected object.
2. the formula slide detection method that contrasts one by one as claimed in claim 1 is characterized in that described contrast material is cultured cell, liquid sample or tissue specimen.
3. the formula slide detection method that contrasts one by one as claimed in claim 2, the cell extraction that it is characterized in that cultured cell is treated to directly and with pancreatin cell is digested from culture flask.
4. the formula slide detection method that contrasts one by one as claimed in claim 2, the cell extraction that it is characterized in that liquid sample is treated to liquid sample and adds that anti-coagulants carries out direct centrifuging, cell is obtained in washing, described cleansing solution is that 0.01mol/L, PH are 7.4 phosphate buffer or TRIS buffer, and anti-coagulants is heparin or citric acid.
5. the formula slide detection method that contrasts one by one as claimed in claim 2 is characterized in that the cell extraction of tissue specimen is treated to tissue specimen flush away bloodstain, grinds, and the 100-400 eye mesh screen filters, and carries out cell punching processing, centrifuging, washing.
6. the formula slide detection method that contrasts one by one as claimed in claim 5 is characterized in that carrying out after stale tissue specimen grinds in time fixing, and immobile liquid is 10-20% neutral formalin or 90-95% ethanol or cold acetone.
7. the formula slide detection method that contrasts one by one as claimed in claim 5, it is characterized in that tissue specimen grinds the back and adds the erythrocyte cracked liquid processing, the preparation of erythrocyte cracked liquid: take by weighing 3-4g ammonium chloride, 1-2g trishydroxymethylaminomethane, be dissolved in water and be diluted to 500ml, 0.22 the degerming of μ m membrane filtration, 4-6 ℃ of preservation.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2005100503073A CN100360925C (en) | 2005-05-12 | 2005-05-12 | Slide detection method employing one-by-one comparison |
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| CNB2005100503073A CN100360925C (en) | 2005-05-12 | 2005-05-12 | Slide detection method employing one-by-one comparison |
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| CN1699953A CN1699953A (en) | 2005-11-23 |
| CN100360925C true CN100360925C (en) | 2008-01-09 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101957282A (en) * | 2010-10-20 | 2011-01-26 | 杭州市中医院 | Preparation method and application of single cell or tissue particle suspension |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104655475A (en) | 2015-02-06 | 2015-05-27 | 丁伟 | Preparation method and application of paraffin embedding contrast suspension |
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| EP0469766A1 (en) * | 1990-07-23 | 1992-02-05 | Becton, Dickinson and Company | Preservation of cells as controls or standards in cellular analysis |
| JPH09121890A (en) * | 1995-10-30 | 1997-05-13 | Mitsubishi Kagaku B C L:Kk | Detection of bacterium belonging to helicobacter |
| CN1356553A (en) * | 2001-09-13 | 2002-07-03 | 陕西超英生物医学研究开发有限公司 | Tissue microarray biochip |
| CN1480732A (en) * | 2003-07-15 | 2004-03-10 | 武汉大学 | Method of nano amplitication detection |
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2005
- 2005-05-12 CN CNB2005100503073A patent/CN100360925C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0469766A1 (en) * | 1990-07-23 | 1992-02-05 | Becton, Dickinson and Company | Preservation of cells as controls or standards in cellular analysis |
| JPH09121890A (en) * | 1995-10-30 | 1997-05-13 | Mitsubishi Kagaku B C L:Kk | Detection of bacterium belonging to helicobacter |
| CN1356553A (en) * | 2001-09-13 | 2002-07-03 | 陕西超英生物医学研究开发有限公司 | Tissue microarray biochip |
| CN1480732A (en) * | 2003-07-15 | 2004-03-10 | 武汉大学 | Method of nano amplitication detection |
Non-Patent Citations (2)
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| 免疫组化对照芯片的设计和应用. 郎志强等.临床与实验病理学杂志，第21卷第2期. 2005 * |
| 用RT-PCR技术检测石蜡包埋组织中的肠道病毒RNA. 李相军等.吉林大学学报（医学版），第28卷第6期. 2002 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101957282A (en) * | 2010-10-20 | 2011-01-26 | 杭州市中医院 | Preparation method and application of single cell or tissue particle suspension |
| CN101957282B (en) * | 2010-10-20 | 2013-06-05 | 杭州市中医院 | Preparation method and application of single cell or tissue particle suspension |
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| CN1699953A (en) | 2005-11-23 |
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Owner name: ZHEJIANG YIBAI BIOLOGY TECHNOLOGY CO., LTD. Free format text: FORMER OWNER: HANGZHOU CHINESE MEDICINAL HOSPITAL Effective date: 20150618 |
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Effective date of registration: 20150618 Address after: 324111, 3, Yu Da Shan village, silted village, Jiangshan Town, Jiangshan, Zhejiang Patentee after: Zhejiang easy Biotechnology Co., Ltd. Address before: 310007 No. 453, Stadium Road, Hangzhou, Zhejiang Patentee before: Hangzhou Chinese Medicinal Hospital |
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