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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: To explore a rapid liver tissue clearing method based on CLARITY technology and provide the basis for the application of this technique on the liver. METHODS: Hydrogels were perfused into rats, and the liver was removed and coagulated to a thickness of 1 mm. The slices were randomly divided into two groups. The control group was treated by the conventional passive lipid clearance method. The experimental group was placed in a special electrophoresis device, and the lipid was washed under the electric field. The optimal liver transparence was explored by increasing the electrophoresis temperature and quantifying the tissue transparency Condition. Results: In the experimental group, the relative transparency of the liver sections reached the highest at 48 h after electrophoresis at 37 ° C for 48 hours, and then the relative transparency began to decrease. After 48 hours of electrophoresis, the temperature was increased and the temperature was maintained for 48 hours. The relative transparency of the liver sections Further increase and the control group lipid clearance rate was significantly lower than the experimental group. Conclusion: The rapid lipid clearance can be achieved when the liver tissue sections are electrophoresed at 37 ° C for 48 hours and then incubated at 52 ° C for 48 hours at 12V.