花鼠脑组织灌流固定方法的探究  

Simple Method for the Perfusion-Fixation of Brain Tissue for Siberian Chipmunk(Tamias sibiricus)

作　　者：毛剑杰 朱青青 陈坤 周洪昌[1] 钱晶[1] 潘永良 MAO Jianjie;ZHU Qingqing;CHEN Kun;ZHOU Hongchang;QIAN Jing;PAN Yongliang(School of Medicine,Huzhou University,Key Laboratory of Vector Biology andPathogen Control of Zhejiang Province,Huzhou,313000,China)

机构地区：[1]湖州师范学院医学院浙江省媒介生物学与病原控制重点实验室

出　　处：《野生动物学报》2019年第3期622-626,共5页CHINESE JOURNAL OF WILDLIFE

基　　金：国家自然科学基金(31672295);湖州师范学院“大学生创新创业训练计划”项目(HUTC-15)

摘　　要：灌流固定过程是影响脑组织结构的关键步骤,关系到后续实验结果的可靠性。目前已有外周组织灌流固定的常规操作步骤,但脑组织耐氧能力差,且缺乏相关的细节描述和效果评估,为提高适用于小型啮齿动物的脑组织灌流固定效果,以花鼠为研究对象,比较了是否主动脉结扎及不同生理盐水温度对脑组织在体心脏灌流固定效果的影响。结扎腹主动脉可减少生理盐水的用量,而对脑组织硬度,器官颜色变化,海马DG区DCX阳性细胞密度没有影响;4℃生理盐水相比于37℃可显著增加海马DG区DCX阳性细胞数,而对脑组织硬度,器官颜色变化没有影响。结果表明,通过心尖入针,穿过左心室到主动脉升部,再结扎腹主动脉及用4℃生理盐水快速清除血液,是啮齿动物在体心脏灌流固定脑组织较为理想的方法。Perfusion and fixation methods are important for the preparation of brain tissue,and can affect the reliability of follow-up experimental results.Although routine procedures are available for perfusion fixation of peripheral tissues,they are not available for brain tissue.To improve the effect of perfusion fixation of brain tissue for small rodents,we examined the effects of temperature ( 4 ℃ and 37 ℃) of normal saline and the ligation of the abdominal aortas working with Siberian Chipmunk.The ligation of the abdominal aortas reduces the volume of normal saline,but had no effect on the hardness of brain tissue and the density of the DCX-labeled cells in the DG.Furthermore,animals perfused with 4℃ normal saline had higher density of DCX-labeled cells in the DG compared to those which were perfused with 37℃ normal saline.This study indicated passing a blunt-tipped perfusion needle through the ventricle into the ascending aorta and using 4 ℃ normal saline to clear away the blood with the ligation of abdominal aorta may be a better method for brain perfusion and fixation in Siberian chipmunk.

关 键 词：花鼠 脑组织 灌流固定 DCX免疫组化 

分 类 号：Q95-331[生物学—动物学]