CN1699953A - One-to-one control slide detection method - Google Patents
One-to-one control slide detection method Download PDFInfo
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- CN1699953A CN1699953A CN 200510050307 CN200510050307A CN1699953A CN 1699953 A CN1699953 A CN 1699953A CN 200510050307 CN200510050307 CN 200510050307 CN 200510050307 A CN200510050307 A CN 200510050307A CN 1699953 A CN1699953 A CN 1699953A
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- cell
- detection method
- slide detection
- contrast
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- 238000001514 detection method Methods 0.000 title claims description 27
- 238000000605 extraction Methods 0.000 claims abstract description 18
- 238000004321 preservation Methods 0.000 claims abstract description 15
- 210000004027 cell Anatomy 0.000 claims description 44
- 239000007788 liquid Substances 0.000 claims description 25
- 239000002872 contrast media Substances 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 10
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 9
- 238000012545 processing Methods 0.000 claims description 9
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 9
- 210000003743 erythrocyte Anatomy 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 6
- 239000003146 anticoagulant agent Substances 0.000 claims description 6
- 229940127219 anticoagulant drug Drugs 0.000 claims description 6
- 210000004748 cultured cell Anatomy 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 239000012188 paraffin wax Substances 0.000 claims description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 3
- 108010010803 Gelatin Proteins 0.000 claims description 3
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 claims description 3
- 108010019160 Pancreatin Proteins 0.000 claims description 3
- 239000007983 Tris buffer Substances 0.000 claims description 3
- 235000019270 ammonium chloride Nutrition 0.000 claims description 3
- 229940098773 bovine serum albumin Drugs 0.000 claims description 3
- 229920000159 gelatin Polymers 0.000 claims description 3
- 239000008273 gelatin Substances 0.000 claims description 3
- 235000019322 gelatine Nutrition 0.000 claims description 3
- 235000011852 gelatine desserts Nutrition 0.000 claims description 3
- 229960002897 heparin Drugs 0.000 claims description 3
- 229920000669 heparin Polymers 0.000 claims description 3
- 238000005374 membrane filtration Methods 0.000 claims description 3
- 230000007935 neutral effect Effects 0.000 claims description 3
- 229940055695 pancreatin Drugs 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- 238000004080 punching Methods 0.000 claims description 3
- 239000000243 solution Substances 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 3
- 238000000034 method Methods 0.000 abstract description 6
- 230000000875 corresponding effect Effects 0.000 abstract 6
- 238000012360 testing method Methods 0.000 description 6
- 239000006285 cell suspension Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 238000007901 in situ hybridization Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a measuring method for one-to-one contrasting slide in the field of medical measuring methodology and technique, which comprises the following steps: choosing corresponding property of measuring object to do cell extraction; preparing cell corresponding preservation: after doing cell extraction to corresponding property, producing cell corresponding preservation in preserving medium with the ratio of cell extraction and preserving medium 1:1.5; when measuring the slide, pointing or defusing cell corresponding preservation into every corresponding slide; measuring, analyzing and gauging them together with measuring object.
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 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.
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.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100503073A CN100360925C (en) | 2005-05-12 | 2005-05-12 | One-to-one control slide detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100503073A CN100360925C (en) | 2005-05-12 | 2005-05-12 | One-to-one control slide detection method |
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| Publication Number | Publication Date |
|---|---|
| CN1699953A true CN1699953A (en) | 2005-11-23 |
| CN100360925C CN100360925C (en) | 2008-01-09 |
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| CNB2005100503073A Expired - Fee Related CN100360925C (en) | 2005-05-12 | 2005-05-12 | One-to-one control slide detection method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016123868A1 (en) * | 2015-02-06 | 2016-08-11 | 丁伟 | Method for preparing liquid-state dropping or coating pathological quality control product, and uses thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IE76319B1 (en) * | 1990-07-23 | 1997-10-22 | Becton Dickinson Co | Preservation of cells as controls or standards in cellular analysis |
| JPH09121890A (en) * | 1995-10-30 | 1997-05-13 | Mitsubishi Kagaku B C L:Kk | Method for detecting Helicobacter bacteria |
| CN1204405C (en) * | 2001-09-13 | 2005-06-01 | 陕西超英生物医学研究开发有限公司 | Tissue microarray biochip |
| CN1214245C (en) * | 2003-07-15 | 2005-08-10 | 武汉大学 | Method of nano amplitication detection |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016123868A1 (en) * | 2015-02-06 | 2016-08-11 | 丁伟 | Method for preparing liquid-state dropping or coating pathological quality control product, and uses thereof |
| US10359345B2 (en) | 2015-02-06 | 2019-07-23 | Wei Ding | Method for preparing liquid-state dripping or coating pathological quality control product and uses thereof |
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| CN100360925C (en) | 2008-01-09 |
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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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Granted publication date: 20080109 Termination date: 20190512 |