CN101551382A - Cell viability detection kit and preparation method and application thereof - Google Patents
Cell viability detection kit and preparation method and application thereof Download PDFInfo
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- CN101551382A CN101551382A CNA2009100265444A CN200910026544A CN101551382A CN 101551382 A CN101551382 A CN 101551382A CN A2009100265444 A CNA2009100265444 A CN A2009100265444A CN 200910026544 A CN200910026544 A CN 200910026544A CN 101551382 A CN101551382 A CN 101551382A
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Abstract
The invention relates to a cell viability detection kit and a preparation method and an application thereof. The invention is applicable to live cell counting, and is a determining method for directly detecting the cell proliferation, and is also a detection method for estimating the cell totoxicity of the drugs, peptide growth factors and other substances, and is applicable to detect the suspended cell and the anchorage-dependent cell cultured in the cell culture plates. The invention consists of a WST-8 mixed solution storage solution, a reaction stop solution and a buffer diluent solution, wherein the WST-8 mixed solution storage solution contains 50mmol/L WST-8 solution, 2mmol/L PMS and 1mmol/L PIPES buffer solution; the reaction stop solution is 10%(W/V) sodium dodecyl sulfate; and the buffer diluent solution is 1mmol/L PIPES solution.
Description
Technical field
What the present invention relates to is a kind of cell viability detection kit and preparation method thereof, application, be used for viable count, directly detect the assay method of cell proliferation, also be the Cytotoxic detection methods of material such as a kind of evaluation medicine, polypeptide growth factor, be applicable to the suspension of cultivating in the Tissue Culture Plate and the detection of attached cell.
Background technology
The classic method that cell proliferation detects is because radioactive contamination, complex operation, sensitivity are low, instrument and equipment requires height, expense costliness etc. former thereby progressively replaced.What be widely used in the cell proliferation detection at present is that the micro-enzyme reaction of tetramethyl azo azoles salt (MTT) is measured.In the sixties in 20th century, scientist finds that MTT can be reduced to by the Intramitochondrial succinate dehydrogenase of living cells and praises product by the bluish violet first moon, and think that it has the existence of reflection living cells and the potential value of propagation, has confirmed that in nineteen eighty-three mtt assay is a kind of objective indicator of correctly estimating cell proliferation and survival condition.Be elected to be the conventional method of screening into external anti-tumor medicine by U.S. NCI.Since the nineties in 20th century, mtt assay with its easy, quick, easy standardization, do not use advantages such as isotope, show one's talent in the quick test method of the multiple medicines of comforming, its drug sensitivity tests guiding clinical treatment cancer of the stomach, colon, the carcinoma of the rectum and tumor in digestive tract are all obtained good result.
The researcher has carried out various experiments improvement to MTT subsequently, the derivant XTT of MTT occurred, can form and praise (Formazan) product by the water-soluble first moon, has omitted thus to add the step that the dissolving of acidifying isopropyl alcohol is praised the first moon in the conventional mtt assay.But XTT method itself is unstable, and relatively indissoluble needs fresh preparation, and the XTT method form to praise product the first moon be orange colour, some yellow metabolins and reagent all can influence testing result in the cultivating system; Therefore design a kind of novel MTT analog MTS again, MTS can be formed by the living cells reduction and praise product by the water miscible first moon under the condition that electron carrier 1-methoxyl-5-toluphenazine dimethyl suflfate (1-Methoxy PMS) exists.To compare tool easy and simple to handle with the XTT method, high specificity, and it is dark-brown that the first moon of formation is praised product, the external factor influence is few during detection; Higher 2-(4-iodobenzene)-3-(4-the nitrobenzene)-5-(2 of sensitivity has appearred again subsequently, 4-two thio phenyls)-2H-tetrazolium salts (WST-1), carrying out equally dehydrogenase reaction under the condition that PMS exists forms and praises product by the water miscible first moon, but WST-1 solution and PMS powder need configuration separately, carry out ratio during use and mix, operate more loaded down with trivial details; Existing market is generally acknowledged no matter be detection sensitivity or the simple and easy degree the best of operation be 2-(2-methoxyl-4-nitrobenzophenone)-3-(4-nitrobenzophenone)-5-(2,4-disulfonic acid benzene)-2h-tetrazolium list sodium salt (WST-8), a kind of novel tetrazolium, the same with the WST-1 action condition, need the existence of PMS.With WST-8 is reagent configuration principal ingredient, occurred numerous cell proliferation detecting kits based on WST-8 both at home and abroad, but ubiquitous problem is 1. a reagent storage terms of validity weak points, causes waste easily; 2. the detection sensitivity tradition of comparing
3The H method of mixing is still waiting to improve; 3. chromogenic reaction does not have cessation reaction liquid to carry out cessation reaction after finishing, and needs to select voluntarily configuration, makes troubles to experiment, causes poor repeatability detection time.
Summary of the invention
The objective of the invention is to weak point at above-mentioned existing product, a kind of cell viability detection kit and preparation method thereof is provided, uses, with WST-8 is the cell proliferation detecting kit of principal ingredient prescription, be applied to viable count, cell proliferation and Cytotoxic detection, can effectively improve its effective service life and detection sensitivity, avoid unnecessary waste.
Cell viability detection kit and preparation method thereof, application are to take following scheme to realize:
Cell viability detection kit is made up of WST-8 mixed solution storage liquid, reaction terminating liquid, buffer diluent.Wherein WST-8 mixed solution storage liquid includes 50mmol/L WST-8 solution, 2mmol/L PMS, 1mol/L PIPES damping fluid; Reaction terminating liquid is 10% (W/V) sodium dodecylsulphonate; Buffer diluent is a 1mol/L PIPES solution.
The preparation method of cell viability detection kit:
(1) configuration of 1mol/L PIPES damping fluid
Take by weighing the free acid of PIPES (piperazine-N, N-two (2-ethanesulfonic acid)) by proportioning, be dissolved in the deionized water, regulate PH to 6.9, add deionized water and be settled to 1mol/L with NaOH, the millipore filter filtration sterilization, 4 ℃ of preservations are standby;
(2) WST-8 mixes the configuration of storage liquid
Take by weighing the WST-8 powder respectively by proportioning, the PMS powder is dissolved in the 1mol/L PIPES damping fluid, mix, and the millipore filter filtration sterilization, 4 ℃ keep in Dark Place;
(3) configuration of reaction terminating liquid
Take by weighing sodium dodecylsulphonate by proportioning and be dissolved in the deionized water, stir, be configured to the sodium dodecyl sulfate solution of 10% (W/V), 4 ℃ of preservations are standby;
(4) assembling process of kit
With the solution of above-mentioned configuration, be sub-packed in the reagent bottle with 10ml, 100ml or the sterile working of 500ml specification, form cell viability detection kit;
Wherein WST-8 mixed solution storage liquid includes 50mmol/L WST-8 solution, 2mmol/L PMS and 1mol/L PIPES damping fluid; Reaction terminating liquid is 10% (W/V) sodium dodecylsulphonate; Buffer diluent is a 1mol/L PIPES solution.
Described WST-8 is commercially available 2-(2-methoxyl-4-nitrobenzophenone)-3-(4-nitrobenzophenone)-5-(2,4-disulfonic acid benzene)-2h-tetrazolium list sodium salt.
Described cell viability detection kit is used for viable count, cell proliferation and Cytotoxic detection.
Adopt PIPES as the solution damping fluid in the kit of the present invention, stable WST-8 reagent reacting condition and being quick on the draw property can be provided, the pKa value of PIPES is near physiological pH, in by cell membrane, can not be absorbed, and can see through ultraviolet ray, can stablize 172 hours at 37 ℃ of PIPES, be better than other damping fluid, often be used to dispose dye liquor and detect tumour cell etc.; The WST-8 mixed solution is configured to storage liquid concentration in addition, for the 5-10 of working concentration doubly, can effectively prolong the Preservation of Reagent time like this; Detect the stable of product for keeping at last, disposed reaction terminating liquid, can effectively control the experiment reaction time, be beneficial to the repeatability of experiment.
Embodiment
Cell viability detection kit is made up of WST-8 mixed solution storage liquid, reaction terminating liquid, buffer diluent.Wherein WST-8 mixed solution storage liquid includes 50mmol/L WST-8 solution, 2mmol/L PMS, 1mol/L PIPES damping fluid; Reaction terminating liquid is 10% (W/V) sodium dodecylsulphonate; Buffer diluent is a 1mol/L PIPES solution.Described WST-8 is commercially available 2-(2-methoxyl-4-nitrobenzophenone)-3-(4-nitrobenzophenone)-5-(2,4-disulfonic acid benzene)-2h-tetrazolium list sodium salt;
Described WST-8 is commercially available 2-(2-methoxyl-4-nitrobenzophenone)-3-(4-nitrobenzophenone)-5-(2,4-disulfonic acid benzene)-2h-tetrazolium list sodium salt.
The preparation method of cell viability detection kit:
Embodiment 1,
Cell viability detection kit 10ml specification, 100 use amount preparation methods:
(1) configuration of 1mol/L PIPES damping fluid
Take by weighing the free acid of 15.12g PIPES (piperazine-N, N-two (2-ethanesulfonic acid)), be dissolved in the 40ml deionized water, regulate PH to 6.9 with NaOH, it is fixed molten to 50ml to add deionized water, the millipore filter filtration sterilization, and 4 ℃ of preservations are standby;
(2) WST-8 mixes the configuration of storage liquid
Take by weighing WST-8 powder 0.06g, PMS powder 0.61268g is dissolved in the 10ml 1mol/LPIPES damping fluid, mix, and the millipore filter filtration sterilization, 4 ℃ keep in Dark Place;
(3) configuration of reaction terminating liquid
Take by weighing the 5g sodium dodecylsulphonate and be dissolved in the 50ml deionized water, stir, be configured to the sodium dodecyl sulfate solution of 10% (W/V), 4 ℃ of preservations are standby.
(4) assembling process of kit
With the solution of above-mentioned configuration, get liquor capacity according to describing below, sterile working is sub-packed in the cell viability detection kit of forming the 10ml specification in the reagent bottle:
1) WST-8 mixed solution storage liquid 1ml:
2) reaction terminating liquid, 10% (W/V) sodium dodecylsulphonate 10ml;
3) 1mol/L PIPES buffer diluent, 10ml
4) dilution bottle is 1.
Embodiment 2,
Cell viability detection kit 100ml specification, 1000 use amount preparation methods:
(1) configuration of 1mol/L PIPES damping fluid
Take by weighing the free acid of 151.2g PIPES (piperazine-N, N-two (2-ethanesulfonic acid)), be dissolved in the 400ml deionized water, regulate PH to 6.9 with NaOH, it is fixed molten to 500ml to add deionized water, the millipore filter filtration sterilization, and 4 ℃ of preservations are standby;
(2) WST-8 mixes the configuration of storage liquid
Take by weighing WST-8 powder 0.6g, PMS powder 6.1268g is dissolved in the 100ml 1mol/LPIPES damping fluid, mix, and the millipore filter filtration sterilization, 4 ℃ keep in Dark Place;
(3) configuration of reaction terminating liquid
Take by weighing the 100g sodium dodecylsulphonate and be dissolved in the 1000ml deionized water, stir, be configured to the sodium dodecyl sulfate solution of 10% (W/V), 4 ℃ of preservations are standby;
(4) assembling process of kit
With the solution of above-mentioned configuration, get liquor capacity according to describing below, sterile working is sub-packed in the cell viability detection kit of forming the 100ml specification in the reagent bottle:
1) WST-8 mixed solution storage liquid 10ml:
2) reaction terminating liquid, 10% (W/V) sodium dodecylsulphonate 100ml;
3) 1mol/L PIPES buffer diluent, 100ml
4) dilution bottle is 1.
Embodiment 3,
Cell viability detection kit 500ml specification, 5000 use amount preparation methods:
(1) configuration of 1mol/L PIPES damping fluid
Take by weighing the free acid of 302.4g PIPES (piperazine-N, N-two (2-ethanesulfonic acid)), be dissolved in the 800ml deionized water, regulate PH to 6.9 with NaOH, it is fixed molten to 1000ml to add deionized water, the millipore filter filtration sterilization, and 4 ℃ of preservations are standby;
(2) WST-8 mixes the configuration of storage liquid
Take by weighing WST-8 powder 3g, PMS powder 30.634g is dissolved in the 500ml 1mol/LPIPES damping fluid, mix, and the millipore filter filtration sterilization, 4 ℃ keep in Dark Place;
(3) configuration of reaction terminating liquid
Take by weighing the 500g sodium dodecylsulphonate and be dissolved in the 5000ml deionized water, stir, be configured to the sodium dodecyl sulfate solution of 10% (W/V), 4 ℃ of preservations are standby;
(4) assembling process of kit
With the solution of above-mentioned configuration, get liquor capacity according to describing below, sterile working is sub-packed in the cell viability detection kit of forming the 500ml specification in the reagent bottle:
1) WST-8 mixed solution storage liquid 50ml:
2) reaction terminating liquid, 10% (W/V) sodium dodecylsulphonate 500ml;
3) 1mol/L PIPES buffer diluent, 500ml
4) dilution bottle is 1.
Described cell viability detection kit is used for viable count, cell proliferation and Cytotoxic detection.
Cell viability detection kit uses:
One, described cell viability detection kit is applied to the detection use flow process of viable count, cell proliferation:
(1) get the growth conditions good cell and be prepared into certain cell concentration suspension, every hole 100 μ l add in the 96 porocyte culture plates, if attached cell needs 37 ℃ of incubators to cultivate 2-4 hour in advance, if suspension cell does not then need pre-cultivation;
(2) with the consumption as required of WST-8 mixed solution storage liquid in the kit, in dilution bottle, use the dilution in 1: 10 of 1mol/L PIPES buffer diluent, standby;
(3) solution in the dilution bottle is added 96 porocyte culture plates according to every hole 10 μ l, 37 ℃ of incubators were cultivated 1-4 hour;
(4) after the chromogenic reaction, add the every hole of 10% (W/V) sodium dodecylsulphonate 10 μ l cessation reactions, 450-490nm photometry absorbance log on the microplate reader.Advise that every increment originally does three repeating holes, will not contain cell, the hole that only adds nutrient culture media and reagent is as the blank hole.But the cell number in the every hole of absorbance OD value indirect reaction in every hole of surveying is an X-axis with one group of cell number gradient, and absorbance OD value is a Y-axis, makes regression curve, can calculate every porocyte number according to the OD value.
Two, described cell viability detection kit is applied to the detection use flow process of thin toxicity alive:
(1) get the growth conditions good cell and be prepared into certain cell concentration suspension, every hole 100 μ l add in the 96 porocyte culture plates, if attached cell needs 37 ℃ of incubators to cultivate 2-4 hour in advance, if suspension cell does not then need pre-cultivation;
(2) toxicant of adding variable concentrations, 37 ℃ of incubators are cultivated, and the time is determined according to the character of toxicant and the susceptibility of cell;
(3) with the consumption as required of WST-8 mixed solution storage liquid in the kit, in dilution bottle, use the dilution in 1: 10 of 1mol/L PIPES buffer diluent, standby;
(4) solution in the dilution bottle is added 96 porocyte culture plates according to every hole 10 μ l, 37 ℃ of incubators were cultivated 1-4 hour;
(5) after the chromogenic reaction, add the every hole of 10% (W/V) sodium dodecylsulphonate 10 μ l cessation reactions, 450-490nm photometry absorbance log on the microplate reader.Advise that every increment originally is three repeated experiments hole As, will not contain cell and toxicant, the hole that only adds nutrient culture media and reagent is as blank hole Ab; Contain the nutrient culture media and the reagent of cell, but the hole that does not add toxicant is as positive control hole Ac.According to the cell survival rate formula: cell survival rate (%)=[(As-Ab)/(Ac-Ab)] * 100% calculates cell survival rate, reflects the cytotoxicity of detected material according to cell survival rate.
Use the present invention with the cytotoxicity that murine myeloma cell SP2/0 detects the HEPES powder below, can be used for the configuration of cell culture medium to determine HEPES.
The murine myeloma cell SP2/0 of complete 1640 medium culture that growth conditions is good, behind the blood counting chamber counting, being prepared into concentration is 1x10
4Cell suspension.Every hole drips 100 μ l cell suspensions in 96 porocyte culture plates, 37 ℃ of pre-cultivations 2 hours of cell culture incubator; Add then in proportion that concentration disposes 10 μ l HEPES solution, 1%DMSO is as the cytotoxicity negative control, and 37 ℃ of cell culture incubators were cultivated 4 hours; Get WST-8 mixed solution storage liquid 100 μ l in the kit, be diluted to 1ml with 900 μ l 1mol/L PIPES in dilution bottle, then the every hole 10 μ l of solution in the dilution bottle are added Tissue Culture Plate, 37 ℃ of incubators were cultivated 3 hours; After observing chromogenic reaction, every hole adds 10 μ l10% (W/V) sodium dodecylsulphonate cessation reactions, and 450nm, 490nm dual wavelength detect absorbance on the microplate reader.Three repetitions are done in each hole, will not contain cell and toxicity sample to be tested HEPES, and the nutrient culture media and the reagent of cell are contained as the blank hole in the hole that only adds nutrient culture media and reagent, but the hole that does not add toxicity sample to be tested HEPES is as the positive control hole.
Experimental result: microscopically is observed and is added in the hole of DMSO as the cytotoxicity negative control, and cell is all dead, and does not add in the positive control hole of toxicity sample to be tested HEPES, and cell growth state is good.Assay kit detects data, and it is 0.051 that negative control hole detects mean value, and blank mean value is 0.021, and positive control mean value is 1.654, and experimental data is consistent with the microexamination result.The HEPES solution that this experiment detects is under variable concentrations, and detecting data readings mean value is 1.398, and the pair cell avirulence can be used for the configuration of biological buffer.
Described PMS adopts commercially available 1-methoxyl-5-toluphenazine dimethyl suflfate (1-Methoxy PMS).
Claims (6)
1, a kind of cell viability detection kit, it is characterized in that being made up of WST-8 mixed solution storage liquid, reaction terminating liquid, buffer diluent, wherein WST-8 mixed solution storage liquid includes 50mmol/L WST-8 solution, 2mmol/L PMS and 1mol/L PIPES damping fluid; Reaction terminating liquid is 10% (W/V) sodium dodecylsulphonate; Buffer diluent is a 1mol/L PIPES solution.
2, the preparation method of the described cell viability detection kit of claim 1 is characterized in that:
(1) configuration of 1mol/L PIPES damping fluid
Take by weighing the free acid of PIPES by proportioning, be dissolved in the deionized water, regulate PH to 6.9, add deionized water and be settled to 1mol/L with NaOH, the millipore filter filtration sterilization, 4 ℃ of preservations are standby;
(2) WST-8 mixes the configuration of storage liquid
Take by weighing the WST-8 powder respectively by proportioning, the PMS powder is dissolved in the 1mol/L PIPES damping fluid, mix, and the millipore filter filtration sterilization, 4 ℃ keep in Dark Place;
(3) configuration of reaction terminating liquid
Take by weighing sodium dodecylsulphonate by proportioning and be dissolved in the deionized water, stir, be configured to the sodium dodecyl sulfate solution of 10% (W/V), 4 ℃ of preservations are standby;
(4) assembling process of kit
With the solution of above-mentioned configuration, be sub-packed in the reagent bottle with 10ml, 100ml or the sterile working of 500ml specification, form cell viability detection kit;
Wherein WST-8 mixed solution storage liquid includes 50mmol/L WST-8 solution, 2mmol/L PMS and 1mol/L PIPES damping fluid; Reaction terminating liquid is 10% (W/V) sodium dodecylsulphonate; Buffer diluent is a 1mol/L PIPES solution.
3, cell viability detection kit according to claim 1 and 2 is characterized in that described WST-8 is 2-(2-methoxyl-4-nitrobenzophenone)-3-(4-nitrobenzophenone)-5-(2,4-disulfonic acid benzene)-2h-tetrazolium list sodium salt.
4, claim 1 or 2 described cell viability detection kits are applied to viable count, cell proliferation and Cytotoxic detection.
5, cell viability detection kit according to claim 4 is applied to the detection use flow process of viable count, cell proliferation:
(1) get the growth conditions good cell and be prepared into certain cell concentration suspension, every hole 100 μ l add in the 96 porocyte culture plates, if attached cell needs 37 ℃ of incubators to cultivate 2-4 hour in advance, if suspension cell does not then need pre-cultivation;
(2) with the consumption as required of WST-8 mixed solution storage liquid in the kit, in dilution bottle, use 1mol/L PIPES to carry out dilution in 1: 10, standby;
(3) solution in the dilution bottle is added 96 porocyte culture plates according to every hole 10 μ l, 37 ℃ of incubators were cultivated 1-4 hour;
(4) after the chromogenic reaction, add the every hole of 10% (W/V) sodium dodecylsulphonate 10 μ l cessation reactions, 450-490nm photometry absorbance log on the microplate reader, every increment is originally done three repeating holes, will not contain cell, the hole that only adds nutrient culture media and reagent is as the blank hole, but the cell number in the every hole of absorbance OD value indirect reaction in every hole of surveying is an X-axis with one group of cell number gradient, and absorbance OD value is a Y-axis, make regression curve, can calculate every porocyte number according to the OD value.
6, cell viability detection kit according to claim 4 is applied to the detection use flow process of thin toxicity alive:
(1) get the growth conditions good cell and be prepared into certain cell concentration suspension, every hole 100 μ l add in the 96 porocyte culture plates, if attached cell needs 37 ℃ of incubators to cultivate 2-4 hour in advance, if suspension cell does not then need pre-cultivation;
(2) toxicant of adding variable concentrations, 37 ℃ of incubators are cultivated, and the time is determined according to the character of toxicant and the susceptibility of cell;
(3) with the consumption as required of WST-8 mixed solution storage liquid in the kit, in dilution bottle, use 1mol/L PIPES to carry out dilution in 1: 10, standby;
(4) solution in the dilution bottle is added 96 porocyte culture plates according to every hole 10 μ l, 37 ℃ of incubators were cultivated 1-4 hour;
(5) after the chromogenic reaction, add the every hole of 10% (W/V) sodium dodecylsulphonate 10 μ l cessation reactions, 450-490nm photometry absorbance log on the microplate reader, every increment originally is three repeated experiments hole As, to not contain cell and toxicant, the hole that only adds nutrient culture media and reagent is as blank hole Ab; Contain the nutrient culture media and the reagent of cell, but the hole that does not add toxicant is as positive control hole Ac; According to the cell survival rate formula: cell survival rate (%)=[(As-Ab)/(Ac-Ab)] x100% calculates cell survival rate, reflects the cytotoxicity of detected material according to cell survival rate.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103834715A (en) * | 2014-03-16 | 2014-06-04 | 国家烟草质量监督检验中心 | WST-8 (water soluble tetrazolium-8) testing method of in vitro cytotoxicity of tobacco juice of electronic cigarette |
| CN106987615A (en) * | 2017-03-21 | 2017-07-28 | 上海恒圆生物科技有限公司 | Cell viability detection reagent and application thereof |
| CN107365827A (en) * | 2016-05-11 | 2017-11-21 | 杭州健昵福生物科技有限公司 | A kind of purposes and method for being used to single sulfonic acid tetrazolium detect cell viability |
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| CN109762871A (en) * | 2017-11-09 | 2019-05-17 | 杭州健昵福生物科技有限公司 | A kind of mixture by single sulfonic acid tetrazolium and PMS derivative is used for the purposes and its detection method of microorganism detection |
| CN113981033A (en) * | 2021-11-16 | 2022-01-28 | 农业农村部环境保护科研监测所 | Method for screening compound with effect of inhibiting cadmium toxicity and/or cadmium absorption and accumulation by using human cells |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103834715A (en) * | 2014-03-16 | 2014-06-04 | 国家烟草质量监督检验中心 | WST-8 (water soluble tetrazolium-8) testing method of in vitro cytotoxicity of tobacco juice of electronic cigarette |
| CN103834715B (en) * | 2014-03-16 | 2015-07-22 | 国家烟草质量监督检验中心 | WST-8 (water soluble tetrazolium-8) testing method of in vitro cytotoxicity of tobacco juice of electronic cigarette |
| CN107365827A (en) * | 2016-05-11 | 2017-11-21 | 杭州健昵福生物科技有限公司 | A kind of purposes and method for being used to single sulfonic acid tetrazolium detect cell viability |
| CN106987615A (en) * | 2017-03-21 | 2017-07-28 | 上海恒圆生物科技有限公司 | Cell viability detection reagent and application thereof |
| CN106987615B (en) * | 2017-03-21 | 2020-07-28 | 上海恒圆生物科技有限公司 | Cell viability detection reagent and application thereof |
| CN109762871A (en) * | 2017-11-09 | 2019-05-17 | 杭州健昵福生物科技有限公司 | A kind of mixture by single sulfonic acid tetrazolium and PMS derivative is used for the purposes and its detection method of microorganism detection |
| CN109536566A (en) * | 2018-11-12 | 2019-03-29 | 北京科技大学 | A kind of method of quick test nano particle to algae activity influence |
| CN109536566B (en) * | 2018-11-12 | 2022-02-25 | 北京科技大学 | Method for rapidly testing influence of nanoparticles on algae activity |
| CN113981033A (en) * | 2021-11-16 | 2022-01-28 | 农业农村部环境保护科研监测所 | Method for screening compound with effect of inhibiting cadmium toxicity and/or cadmium absorption and accumulation by using human cells |
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Application publication date: 20091007 |