CN111077003A - 细胞活性检测微毒性染色剂 - Google Patents
细胞活性检测微毒性染色剂 Download PDFInfo
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- 230000000694 effects Effects 0.000 title claims abstract description 27
- 238000010186 staining Methods 0.000 title claims abstract description 25
- 238000001514 detection method Methods 0.000 title claims abstract description 6
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims abstract description 34
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 26
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- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 claims abstract description 13
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- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims abstract description 13
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims abstract description 13
- 235000010323 ascorbic acid Nutrition 0.000 claims abstract description 13
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- 239000008367 deionised water Substances 0.000 claims abstract description 13
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 13
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 13
- 239000004310 lactic acid Substances 0.000 claims abstract description 13
- 229960000907 methylthioninium chloride Drugs 0.000 claims abstract description 13
- 229910052938 sodium sulfate Inorganic materials 0.000 claims abstract description 13
- 235000011152 sodium sulphate Nutrition 0.000 claims abstract description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 150000002224 folic acids Chemical class 0.000 claims abstract description 9
- 231100000331 toxic Toxicity 0.000 claims description 15
- 230000002588 toxic effect Effects 0.000 claims description 15
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims description 14
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 8
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 claims description 7
- 229960000304 folic acid Drugs 0.000 claims description 7
- 235000019152 folic acid Nutrition 0.000 claims description 7
- 239000011724 folic acid Substances 0.000 claims description 7
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims description 4
- 229930003268 Vitamin C Natural products 0.000 claims description 4
- 229960000583 acetic acid Drugs 0.000 claims description 4
- 239000012362 glacial acetic acid Substances 0.000 claims description 4
- 235000019154 vitamin C Nutrition 0.000 claims description 4
- 239000011718 vitamin C Substances 0.000 claims description 4
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 2
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 claims description 2
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- 231100000053 low toxicity Toxicity 0.000 abstract description 2
- 238000007447 staining method Methods 0.000 description 4
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- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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Abstract
本发明公开了一种细胞活性检测微毒性染色剂,包括以下组分和配比:乳酸4‑8 wt%;亚甲基蓝0.05‑3 wt%;硫酸钠3‑8 wt%;叶酸衍生物16‑18 wt%;抗坏血酸0.5‑7.5 wt%;果糖3‑15 wt%;余量为去离子水。本发明的细胞活性检测微毒性染色剂毒性非常的小,不会影响操作者的健康,成本低且染色效果好,充分满足了生物实验的需要,具有广阔的应用前景。
Description
技术领域
本发明涉及生物技术领域,具体是一种细胞活性检测微毒性染色剂。
背景技术
细胞活性是判断体外培养细胞在例如药物处理、放射性或紫外线照射、培养条件变化等一些条件下是否能正常生长的重要指标。目前常用的细胞活性检测方法有很多,如有染色法,克隆(集落)形成法、比色法、放射性同位素掺入法等。
染色法是细胞培养中检查细胞死活最常用的方法,分为化学染色法和荧光染色法,即利用死细胞和活细胞对染料不同的亲和力,检查细胞活性,染色后在光学或荧光显微镜下即可观察到结果。但化学染色法如若染色时间太长,活细胞也会被染色,干扰对结果的判定,而且化学染色过程中细胞没有被固定,形态也不清晰。而荧光染色法的荧光染料一般都有一定的毒性,影响操作者健康。
发明内容
本发明就是为了解决上述技术问题,所提供了一种细胞活性检测微毒性染色剂。
本发明是按照以下技术方案实现的。
一种细胞活性检测微毒性染色剂,包括以下组分:乳酸、亚甲基蓝、硫酸钠、叶酸衍生物、抗坏血酸、果糖、余量为去离子水。
进一步的,一种细胞活性检测微毒性染色剂,包括以下组分和配比:
乳酸 4-8 wt%;
亚甲基蓝 0.05-3 wt %;
硫酸钠 3-8 wt %;
叶酸衍生物 16-18 wt%;
抗坏血酸 0.5-7.5 wt %;
果糖 3-15 wt%;
余量为去离子水。
进一步的,一种细胞活性检测微毒性染色剂,包括以下组分和配比:
乳酸 6 wt%;
亚甲基蓝 2 wt %;
硫酸钠 5 wt %;
叶酸衍生物 17 wt%;
抗坏血酸 3.5 wt %;
果糖 9 wt%;
余量为去离子水。
进一步的,所述叶酸衍生物为组氨酸 N-乙酰-2-乙二酰氨叶酸。
进一步的,所述抗坏血酸替换为维生素C。
进一步的,所述乳酸替换为冰醋酸。
本发明获得了如下的有益效果。
本发明的细胞活性检测微毒性染色剂毒性非常的小,不会影响操作者的健康,成本低且染色效果好,充分满足了生物实验的需要,具有广阔的应用前景。
附图说明
图1是本发明实施例3对微载体上细胞的染色结果图。
具体实施方式
下面结合附图及实施例对本发明进行进一步说明。
实施例1
一种细胞活性检测微毒性染色剂,包括以下组分和配比:
乳酸 4 wt%;
亚甲基蓝 0.05 wt %;
硫酸钠 3 wt %;
组氨酸 N-乙酰-2-乙二酰氨叶酸 16 wt%;
抗坏血酸 0.5 wt %;
果糖 3 wt%;
余量为去离子水。
实施例2
一种细胞活性检测微毒性染色剂,包括以下组分和配比:
乳酸 8 wt%;
亚甲基蓝 3 wt %;
硫酸钠 8 wt %;
组氨酸 N-乙酰-2-乙二酰氨叶酸 18 wt%;
抗坏血酸 7.5 wt %;
果糖 15 wt%;
余量为去离子水。
实施例3
一种细胞活性检测微毒性染色剂,包括以下组分和配比:
乳酸 6 wt%;
亚甲基蓝 2 wt %;
硫酸钠 5 wt %;
组氨酸 N-乙酰-2-乙二酰氨叶酸 17 wt%;
抗坏血酸 3.5 wt %;
果糖 9 wt%;
余量为去离子水。
按照实施例3各组分和配比配置染色剂,并对微载体上的细胞进行染色,染色结果参见图1.
实施例4
一种细胞活性检测微毒性染色剂,包括以下组分和配比:
乳酸 6 wt%;
亚甲基蓝 2 wt %;
硫酸钠 5 wt %;
组氨酸 N-乙酰-2-乙二酰氨叶酸 17 wt%;
维生素C 3.5 wt %;
果糖 9 wt%;
余量为去离子水。
实施例5
一种细胞活性检测微毒性染色剂,包括以下组分和配比:
冰醋酸 6 wt%;
亚甲基蓝 2 wt %;
硫酸钠 5 wt %;
组氨酸 N-乙酰-2-乙二酰氨叶酸 17 wt%;
抗坏血酸 3.5 wt %;
果糖 9 wt%;
余量为去离子水。
实施例6
一种细胞活性检测微毒性染色剂,包括以下组分和配比:
冰醋酸 6 wt%;
亚甲基蓝 2 wt %;
硫酸钠 5 wt %;
组氨酸 N-乙酰-2-乙二酰氨叶酸 17 wt%;
维生素C 3.5 wt %;
果糖 9 wt%;
余量为去离子水。
此外,应当理解,虽然本说明书按照实施方式加以描述,但并非每个实施方式仅包含一个独立的技术方案,说明书的这种叙述方式仅仅是为清楚起见,本领域技术人员应当将说明书作为一个整体,各实施例中的技术方案也可以经适当组合,形成本领域技术人员可以理解的其他实施方式。
Claims (6)
1.一种细胞活性检测微毒性染色剂,其特征在于:包括以下组分:乳酸、亚甲基蓝、硫酸钠、叶酸衍生物、抗坏血酸、果糖、余量为去离子水。
2.根据权利要求1所述的一种细胞活性检测微毒性染色剂,其特征在于:所述组分的配比为:
乳酸 4-8 wt%;
亚甲基蓝 0.05-3 wt %;
硫酸钠 3-8 wt %;
叶酸衍生物 16-18 wt%;
抗坏血酸 0.5-7.5 wt %;
果糖 3-15 wt%;
余量为去离子水。
3.根据权利要求1所述的一种细胞活性检测微毒性染色剂,其特征在于:所述组分的配比为:
乳酸 6 wt%;
亚甲基蓝 2 wt %;
硫酸钠 5 wt %;
叶酸衍生物 17 wt%;
抗坏血酸 3.5 wt %;
果糖 9 wt%;
余量为去离子水。
4.根据权利要求1所述的一种细胞活性检测微毒性染色剂,其特征在于:所述叶酸衍生物为组氨酸 N-乙酰-2-乙二酰氨叶酸。
5.根据权利要求1所述的一种细胞活性检测微毒性染色剂,其特征在于:所述抗坏血酸替换为维生素C。
6.根据权利要求1所述的一种细胞活性检测微毒性染色剂,其特征在于:所述乳酸替换为冰醋酸。
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103483872A (zh) * | 2013-09-30 | 2014-01-01 | 青岛农业大学 | 用于肿瘤组织细胞的染色剂组合物及其制备方法 |
CN105199430A (zh) * | 2015-11-05 | 2015-12-30 | 济宁博联生物科技有限公司 | 一种上皮组织染色剂及其制备方法 |
CN105571922A (zh) * | 2015-12-10 | 2016-05-11 | 浙江检康生物技术股份有限公司 | 一种上皮组织病变细胞特殊染色试剂及制备工艺 |
CN107238524A (zh) * | 2016-03-29 | 2017-10-10 | 安徽深蓝医疗科技股份有限公司 | 一种改良型特殊染色诊断液的组分及其制备方法 |
CN108318478A (zh) * | 2017-01-18 | 2018-07-24 | 四川天莆高科医疗器械有限公司 | 一种人体上皮组织癌细胞的检测试剂及其制备方法和用途 |
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Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103483872A (zh) * | 2013-09-30 | 2014-01-01 | 青岛农业大学 | 用于肿瘤组织细胞的染色剂组合物及其制备方法 |
CN105199430A (zh) * | 2015-11-05 | 2015-12-30 | 济宁博联生物科技有限公司 | 一种上皮组织染色剂及其制备方法 |
CN105571922A (zh) * | 2015-12-10 | 2016-05-11 | 浙江检康生物技术股份有限公司 | 一种上皮组织病变细胞特殊染色试剂及制备工艺 |
CN107238524A (zh) * | 2016-03-29 | 2017-10-10 | 安徽深蓝医疗科技股份有限公司 | 一种改良型特殊染色诊断液的组分及其制备方法 |
CN108318478A (zh) * | 2017-01-18 | 2018-07-24 | 四川天莆高科医疗器械有限公司 | 一种人体上皮组织癌细胞的检测试剂及其制备方法和用途 |
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