SDS与Triton X-100对细胞膜通透作用研究  

Comparative study on cell membrane permeability of SDS and Triton X-100

作　　者：刘霞 孔华庭[1,2,3] 张继超 诸颖[1,2,3] LIU Xia;KONG Huating;ZHANG Jichao;ZHU Ying(Division of Physical Biology,CAS Key Laboratory of Interfacial Physics and Technology,Shanghai Institute of Applied Physics,Chinese Academy of Sciences,Shanghai 201800,China;The Interdisciplinary Research Center,Shanghai Synchrotron Radiation Facility,Zhangjiang Laboratory,Shanghai Advanced Research Institute,Chinese Academy of Sciences,Shanghai 201210,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院上海应用物理研究所中国科学院微观界面物理与探测重点实验室,上海201800 [2]中国科学院上海高等研究院基础交叉研究中心张江实验室上海光源,上海201210 [3]中国科学院大学,北京100049

出　　处：《核技术》2022年第8期47-52,共6页Nuclear Techniques

基　　金：国家自然科学基金(No.22022410、No.82050005、No.12005281);上海市科委仪器项目(No.19142202400);中国科学院青年创新促进会人才专项(No.2016236)资助。

摘　　要：十二烷基硫酸钠(Sodium Dodecyl Sulfate,SDS)和聚乙二醇辛基苯基醚(Triton X-100)是实验室中常用的表面活性剂,能溶解细胞膜中的脂类物质,增加细胞膜的通透性,辅助染色试剂快速渗透,在免疫染色或组织透明化中发挥着重要的作用。本工作研究了SDS和Triton X-100对细胞膜通透程度的差异,优化了最佳的透膜条件;考察了SDS和Triton X-100的选择对具体实验结果的影响,并应用合肥光源软X射线纳米显微线站的全场透射成像技术研究了透膜前后细胞形态的变化。结果表明:0.25%Triton X-100的透膜效果优于4%SDS。0.25%Triton X-100的最优处理时间为10 min;在过氧化物酶催化3,3’-二氨基联苯胺(3,3’-Diaminobenzidine,DAB)聚合的实验中,SDS会影响酶的催化活性进而影响染色效果,而Triton X-100不会影响染色结果;同步辐射高分辨成像结果显示,采用Triton X-100进行透膜,还不会显著影响细胞的形态。[Background]Sodium dodecyl sulfate(SDS)and polyethylene glycol octyl phenyl ether(Triton X-100)are commonly used for surfactants in the laboratory,which can dissolve lipids in the cell membrane,increase the permeability of cell membrane,and can assist the rapid penetration of staining reagents.Surfactants play an important role in immunostaining or tissue clearance.[Purpose]This study aims to study the effect of SDS and Triton X-100 on cell membrane permeability.[Methods]Firstly,the damage effects of SDS and Triton X-100 on cell membranes were compared and the optimal conditions were optimized.Then,the influence of the selection of SDS and Triton X-100on the specific experimental results was investigated.Finally,the full-field transmission imaging technology of the soft X-ray nanoimaging beamline of the National Synchrotron Radiation Laboratory at Hefei was used to compare their affection on cell morphology before and after the cell rupture.[Results]Experimental results show that 0.25%Triton X-100 has better membrane penetration than 4%SDS,and the optimal treatment time of 0.25%Triton X-100is 10 min.In the experiment of peroxidase catalyzed 3,3’-Diaminobenzidine(DAB)polymerization,SDS,rather than Triton X-100,affects the catalytic activity of enzymes,hence changes the staining effect.The synchrotron radiation high-resolution imaging demonstrate that there is no significant difference of cell morphology before and after the cell membrane permeability with Triton X-100.[Conclusions]Triton X-100 is a better surfactant than SDS in the cell membrane permeability.The principal method of this study provides a good support for selecting suitable surfactants in the experiments.

关 键 词：十二烷基硫酸钠 聚乙二醇辛基苯基醚 细胞膜通透 X射线成像 

分 类 号：TL99[核科学技术—核技术及应用]