CN106769807A - 一种利用流式细胞仪检测HeLa细胞凋亡的方法 - Google Patents
一种利用流式细胞仪检测HeLa细胞凋亡的方法 Download PDFInfo
- Publication number
- CN106769807A CN106769807A CN201611114043.8A CN201611114043A CN106769807A CN 106769807 A CN106769807 A CN 106769807A CN 201611114043 A CN201611114043 A CN 201611114043A CN 106769807 A CN106769807 A CN 106769807A
- Authority
- CN
- China
- Prior art keywords
- hours
- cell
- compound
- hela
- vehicle compounds
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 230000006907 apoptotic process Effects 0.000 title claims abstract description 12
- 238000000034 method Methods 0.000 title claims abstract description 12
- 150000001875 compounds Chemical class 0.000 claims abstract description 31
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims abstract description 14
- -1 poly butylene succinate Polymers 0.000 claims abstract description 11
- 229910002092 carbon dioxide Inorganic materials 0.000 claims abstract description 7
- 239000001569 carbon dioxide Substances 0.000 claims abstract description 7
- 238000004113 cell culture Methods 0.000 claims abstract description 7
- 230000005284 excitation Effects 0.000 claims abstract description 7
- 238000000684 flow cytometry Methods 0.000 claims abstract description 7
- 239000013642 negative control Substances 0.000 claims abstract description 7
- 229920002961 polybutylene succinate Polymers 0.000 claims abstract description 6
- 239000004631 polybutylene succinate Substances 0.000 claims abstract description 6
- 239000003814 drug Substances 0.000 claims description 2
- 229940079593 drug Drugs 0.000 claims description 2
- 238000011275 oncology therapy Methods 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- 238000012360 testing method Methods 0.000 claims description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 1
- ICIWUVCWSCSTAQ-UHFFFAOYSA-M iodate Chemical compound [O-]I(=O)=O ICIWUVCWSCSTAQ-UHFFFAOYSA-M 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
Landscapes
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
一种利用流式细胞仪检测 HeLa细胞凋亡的方法,包括:步骤1:将处于对数生长期HeLa细胞按照100000个细胞/孔分于6孔培养板中,置于37℃、5%二氧化碳,相对湿度为95%的细胞培养箱培养24小时;步骤2:加入不同浓度的第一化合物,浓度分别是:2.5、5和10μM,并以0.1%的溶媒化合物作为阴性对照组,孵育24小时;步骤3:用不含溶媒化合物的胰酶消化收集,用聚丁二酸丁二醇酯洗涤细胞二次;步骤6:1小时内进行流式细胞仪分析,激发波长Ex=488nm,发射波长Em=530nm。
Description
技术领域
本发明涉及一种测试方法,尤其是一种利用流式细胞仪检测 HeLa细胞凋亡的方法。
背景技术
现有技术中对HeLa细胞凋亡的测试精度不够,方法繁琐,无法低成本推广。
本申请的发明人通过潜心研究找到一种高效,高精度测量HeLa细胞凋亡的方法。
发明内容
本发明为了提供一种利用流式细胞仪检测 HeLa细胞凋亡的方法,包括:步骤1:将处于对数生长期HeLa细胞按照100000个细胞/孔分于6孔培养板中,置于37℃、5%二氧化碳,相对湿度为95%的细胞培养箱培养24小时;步骤2:加入不同浓度的第一化合物,浓度分别是:2.5、5和10μM,并以0.1%的溶媒化合物作为阴性对照组,孵育24小时;第一化合物的分子结构式是:;溶媒化合物的分子结构式是:
;步骤3:用不含溶媒化合物的胰酶消化收集,用聚丁二酸丁二醇酯洗涤细胞二次,(2000rpm离心5min),收集100000-200000个细胞;步骤4:加入5μL碘化丙啶,混匀;步骤5:室温、避光、反应5分钟;步骤6:1小时内进行流式细胞仪分析,激发波长Ex=488nm,发射波长Em=530nm。
本发明可以广泛应用到抗癌药物的测试中。
具体实施方式
实施例1:
一种利用流式细胞仪检测 HeLa细胞凋亡的方法,包括:
步骤1:将处于对数生长期HeLa细胞按照100000个细胞/孔分于6孔培养板中,置于37℃、5%二氧化碳,相对湿度为95%的细胞培养箱培养24小时;
步骤2:加入不同浓度的第一化合物,浓度分别是:25、50和100μM,并以0.1%的溶媒化合物作为阴性对照组,孵育24小时;第一化合物的分子结构式是:
;
溶媒化合物的分子结构式是:
步骤3:用不含溶媒化合物的胰酶消化收集,用聚丁二酸丁二醇酯洗涤细胞二次,(2000rpm离心5min),收集100000-200000个细胞;
步骤4:加入5μL碘化丙啶,混匀;
步骤5:室温、避光、反应5分钟;-
步骤6:1小时内进行流式细胞仪分析,激发波长Ex=488nm,发射波长Em=530nm。
实施例2:
一种利用流式细胞仪检测 HeLa细胞凋亡的方法,包括:
步骤1:将处于对数生长期HeLa细胞按照100000个细胞/孔分于6孔培养板中,置于37℃、5%二氧化碳,相对湿度为95%的细胞培养箱培养24小时;
步骤2:加入不同浓度的第一化合物,浓度分别是:0.5、1和2μM,并以0.1%的溶媒化合物作为阴性对照组,孵育24小时;第一化合物的分子结构式是:
;
溶媒化合物的分子结构式是:
步骤3:用不含溶媒化合物的胰酶消化收集,用聚丁二酸丁二醇酯洗涤细胞二次,(2000rpm离心5min),收集100000-200000个细胞;
步骤4:加入5μL碘化丙啶,混匀;
步骤5:室温、避光、反应5分钟;-
步骤6:1小时内进行流式细胞仪分析,激发波长Ex=488nm,发射波长Em=530nm。
实施例3:
一种利用流式细胞仪检测 HeLa细胞凋亡的方法,包括:
步骤1:将处于对数生长期HeLa细胞按照100000个细胞/孔分于6孔培养板中,置于37℃、5%二氧化碳,相对湿度为95%的细胞培养箱培养24小时;
步骤2:加入不同浓度的第一化合物,浓度分别是:1、2和4μM,并以0.1%的溶媒化合物作为阴性对照组,孵育24小时;第一化合物的分子结构式是:
;
溶媒化合物的分子结构式是:
步骤3:用不含溶媒化合物的胰酶消化收集,用聚丁二酸丁二醇酯洗涤细胞二次,(2000rpm离心5min),收集100000-200000个细胞;
步骤4:加入5μL碘化丙啶,混匀;
步骤5:室温、避光、反应5分钟;-
步骤6:1小时内进行流式细胞仪分析,激发波长Ex=488nm,发射波长Em=530nm。
实施例4:
一种利用流式细胞仪检测 HeLa细胞凋亡的方法,包括:
步骤1:将处于对数生长期HeLa细胞按照100000个细胞/孔分于6孔培养板中,置于37℃、5%二氧化碳,相对湿度为95%的细胞培养箱培养24小时;
步骤2:加入不同浓度的第一化合物,浓度分别是:5、10和20μM,并以0.1%的溶媒化合物作为阴性对照组,孵育24小时;第一化合物的分子结构式是:
;
溶媒化合物的分子结构式是:
步骤3:用不含溶媒化合物的胰酶消化收集,用聚丁二酸丁二醇酯洗涤细胞二次,(2000rpm离心5min),收集100000-200000个细胞;
步骤4:加入5μL碘化丙啶,混匀;
步骤5:室温、避光、反应5分钟;-
步骤6:1小时内进行流式细胞仪分析,激发波长Ex=488nm,发射波长Em=530nm。
Claims (2)
1.一种利用流式细胞仪检测 HeLa细胞凋亡的方法,包括:
步骤1:将处于对数生长期HeLa细胞按照100000个细胞/孔分于6孔培养板中,置于37℃、5%二氧化碳,相对湿度为95%的细胞培养箱培养24小时;
步骤2:加入不同浓度的第一化合物,浓度分别是:25、50和100μM,并以0.1%的溶媒化合物作为阴性对照组,孵育24小时;第一化合物的分子结构式是:
溶媒化合物的分子结构式是:
步骤3:用不含溶媒化合物的胰酶消化收集,用聚丁二酸丁二醇酯洗涤细胞二次,(2000rpm离心5min),收集100000-200000个细胞;
步骤4:加入5μL碘化丙啶,混匀;
步骤5:室温、避光、反应5分钟;-
步骤6:1小时内进行流式细胞仪分析,激发波长Ex=488nm,发射波长Em=530nm。
2.本发明可以广泛应用到抗癌药物的测试中。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201611114043.8A CN106769807A (zh) | 2016-12-07 | 2016-12-07 | 一种利用流式细胞仪检测HeLa细胞凋亡的方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201611114043.8A CN106769807A (zh) | 2016-12-07 | 2016-12-07 | 一种利用流式细胞仪检测HeLa细胞凋亡的方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN106769807A true CN106769807A (zh) | 2017-05-31 |
Family
ID=58874665
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201611114043.8A Pending CN106769807A (zh) | 2016-12-07 | 2016-12-07 | 一种利用流式细胞仪检测HeLa细胞凋亡的方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN106769807A (zh) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105358177A (zh) * | 2013-04-17 | 2016-02-24 | 西格诺药品有限公司 | 包含tor激酶抑制剂和imid化合物的联合疗法用于治疗癌症 |
CN105793255A (zh) * | 2013-10-04 | 2016-07-20 | 无限药品股份有限公司 | 杂环化合物及其用途 |
CN105916882A (zh) * | 2013-12-16 | 2016-08-31 | 得克萨斯技术大学联合体 | 作为用于靶向癌症治疗的细胞毒性药物递送系统的抗-ron单克隆抗体 |
-
2016
- 2016-12-07 CN CN201611114043.8A patent/CN106769807A/zh active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105358177A (zh) * | 2013-04-17 | 2016-02-24 | 西格诺药品有限公司 | 包含tor激酶抑制剂和imid化合物的联合疗法用于治疗癌症 |
CN105793255A (zh) * | 2013-10-04 | 2016-07-20 | 无限药品股份有限公司 | 杂环化合物及其用途 |
CN105916882A (zh) * | 2013-12-16 | 2016-08-31 | 得克萨斯技术大学联合体 | 作为用于靶向癌症治疗的细胞毒性药物递送系统的抗-ron单克隆抗体 |
Non-Patent Citations (1)
Title |
---|
李丹等: "苦参碱对宫颈癌HeLa细胞的作用", 《武汉大学学报(医学版)》 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Chen et al. | Reagents-loaded, automated assay that integrates recombinase-aided amplification and Cas12a nucleic acid detection for a point-of-care test | |
Shi et al. | Ultrasensitive and facile detection of microRNA via a portable pressure meter | |
Cheong et al. | Fast detection of SARS-CoV-2 RNA via the integration of plasmonic thermocycling and fluorescence detection in a portable device | |
Deng et al. | Rapid one-step detection of viral particles using an aptamer-based thermophoretic assay | |
Ma et al. | Copper-mediated DNA-scaffolded silver nanocluster on–off switch for detection of pyrophosphate and alkaline phosphatase | |
Ouyang et al. | Universal amplification-free molecular diagnostics by billion-fold hierarchical nanofluidic concentration | |
Zhou et al. | High-fidelity CRISPR/Cas13a trans-cleavage-triggered rolling circle amplified DNAzyme for visual profiling of microRNA | |
Hu et al. | A 2D bilayered metal–organic framework as a fluorescent sensor for highly selective sensing of nitro explosives | |
Bej et al. | “Naked-eye” detection of CN− from aqueous phase and other extracellular matrices: an experimental and theoretical approach mimicking the logic gate concept | |
Chen et al. | 2D MOF nanosensor‐integrated digital droplet microfluidic flow cytometry for in situ detection of multiple miRNAs in single CTC cells | |
Zhao et al. | An ideal detector composed of a 3D Gd-based coordination polymer for DNA and Hg 2+ ion | |
CN105004775A (zh) | 二硫化物点/纳米片复合物dna电化学探针的制备方法 | |
CN103115903B (zh) | 一种微量四环素类抗生素的荧光检测方法 | |
Zong et al. | Automated centrifugal microfluidic chip integrating pretreatment and molecular diagnosis for hepatitis B virus genotyping from whole blood | |
Li et al. | A label-free conjugated polymer-based fluorescence assay for the determination of adenosine triphosphate and alkaline phosphatase | |
Zhou et al. | Chemiluminescence sensor for miRNA-21 detection based on CRISPR-Cas12a and cation exchange reaction | |
Hu et al. | Cluster-based metal–organic frameworks as sensitive and selective luminescent probes for sensing nitro explosives | |
Jiang et al. | A Novel d‐f Heterometallic CdII‐EuIII Metal‐organic Framework as a Sensitive Luminescent Sensor for the Dual Detection of Ronidazole and 4‐Nitrophenol | |
CN103992788A (zh) | 六苯并苯衍生物探针、制备方法及基于六苯并苯衍生物探针与核酸适配体的蛋白质检测方法 | |
Zhou et al. | [2] Pseudorotaxane‐Based Supramolecular Optical Indicator for the Visual Detection of Cellular Cyanide Excretion | |
Li et al. | Multiplex nucleic acid assay of SARS-CoV-2 via a lanthanide nanoparticle-tagging strategy | |
Kang et al. | Fluorescent chemosensor based on bispicolylamine for selective detection of magnesium ions | |
Ma et al. | Flap endonuclease-induced steric hindrance change enables the construction of multiplex and versatile lateral flow strips for DNA detection | |
CN106769807A (zh) | 一种利用流式细胞仪检测HeLa细胞凋亡的方法 | |
Tierno et al. | Next-generation sequencing and triple-negative breast cancer: insights and applications |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WD01 | Invention patent application deemed withdrawn after publication | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170531 |