基于电泳温度提升建立大鼠肝快速透明化模型  

Elevated Temperature Induced Acceleration of Electrophoretic Tissue Clearing Process of Rat Liver Tissue

作　　者：袁小川 连芳[2] 赵荫农[1] 陈闯[1] 黄山[1] 李科志[1] 曾爱屏[1] 何剑波[1] 邬国斌[1] YUAN Xiao-chuan LIAN Fang ZHAO Yin-nong CHEN Chuang HUANG Shan LI Ke-zhi ZENG Ai-ping HE Jian-bo WU Guo-bin(Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China Department of Physiology, Basic Medical School of Guangxi Medical University, Nanning 530021, China)

机构地区：[1]广西医科大学附属肿瘤医院,南宁530021 [2]广西医科大学基础医学院,南宁530021

出　　处：《中国生物工程杂志》2017年第9期65-70,共6页China Biotechnology

基　　金：国家自然科学基金(81460426);广西壮族自治区医药卫生适宜技术推广项目(S201630)资助项目

摘　　要：目的:探索一种基于CLARITY技术的快速肝组织透明化手段,为该技术在肝脏上的应用提供研究基础。方法:水凝胶灌注大鼠,取出肝脏待水凝胶凝固后切为1mm薄片。切片随机分为两组,对照组采用常规被动脂质清除法处理,实验组置于特制电泳装置中,在外加电场条件下洗涤脂质,通过提高电泳温度并量化组织透明度,探索最优肝透明化条件。结果:实验组在12V电压下37℃电泳48h时肝切片相对透明度达到最高并保持至96h,随后相对透明度开始下降,而在48h电泳清除脂质后提高温度继续电泳48h,肝组织切片相对透明度可进一步提高且对照组脂质清除速率明显低于实验组。结论:在12V电压下肝组织切片37℃电泳48h再辅以52℃电泳48h可以实现较快速的脂质清除。Objective： CLARIT is a morphological technique that produces structurally intact,yet optically transparent tissue,which has important applications in nervous system structure and function investigations.However in other organs,especially liver tissue,the researches are not comprehensive. Since the lipid in liver tissue is difficult to clear. The protocol is aim to make this process faster and easier. Method： The rat livers are fixed with hydrogel and cut into 1 mm slices. The control group is cleared with passive method. On the other side,the experimental group is dealt with electrophoretic tissue clearing and elevate temperature as well to find the optimum temperature for the liver tissue clearing. Results： Under the condition of 12 V 37℃,the experimental group become transparent after 48 h electrophoresis,the relative-transparency is changeless when run with another 48 h electrophoresis. If elevate the temperature of electrophoresis,after 48 h basal electrophoresis（ 12 V 37℃）,the relative-transparency can be further improved. Conclusion： To create extremely clear liver tissue,the best electrophoretic tissue clearing procedure is to run the samples at 12 V 37℃ for 48 h followed by more 48 h with the temperature increased to 52℃.

关 键 词：CLARITY 电泳温度 肝切片 脂质清除 相对透明度 

分 类 号：Q819[生物学—生物工程]