大鼠脑组织灌注固定方法的改进  

A modified method for perfusion fixation of rat brain tissue

作　　者：卢文朋 栗世方[1] 

机构地区：[1]青岛大学附属医院神经外科,青岛医学硕士研究生266003

出　　处：《医学研究生学报》2014年第6期627-629,共3页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81070942)

摘　　要：目的免疫荧光等实验研究中良好的组织灌注固定效果对实验结果至关重要。改进大鼠脑组织灌注固定操作方法,提高灌注固定效果。方法改变以往开胸术式,沿腹中线自剑突剪开皮肤至下颌部,暴露两侧胸廓,游离胸前壁;处理灌注针头,剪除针尖,左手捏持心脏,右手持眼科剪在心尖部剪开一小口,将灌注针沿小口插入左心室并推送至升主动脉,固定使之不能退出;夹闭胸主动脉后灌注固定,打开调节阀,灌注等渗盐水,剪开右心耳,当流出液体清亮时,不拔出灌注针,更换输液管,4%多聚甲醛进行固定,先快后慢,颈部僵硬后,完成固定。结果采用此方法,大鼠只有头颈部和双上肢完成固定,较传统方法省时、省液,免疫荧光图片清晰。结论改进后的操作,提高了灌注固定效果,值得推广。Objective Effective tissue perfusion fixation plays an important role in immunofluorescence experiments and other studies.The aim of this study was to modify the method for perfusion fixation of rat brain tissues and improve the efficiency of the procedure.Methods Conventional thoracotomy was modified by cutting the skin from the xiphoid along the mediventral line to the lower jaw to expose both sides of the thorax and then freeing the chest wall.The tip of the infusion needle was cut off and the needle inserted into the left ventricle,pushed to the ascending aorta,and fixed there to prevent sliding.Perfusion and fixation began after thoracic aortic occlusion.The regulatory valve was opened,followed by infusion of 0.9% physiological saline and cutting the right atrial appendage.When the perfusate became clear,the infusion tube was replaced,without removal of the needle,and fixed with 4%paraformaldehyde.The fixation process was slowed down gradually till the neck became stiff,which indicated its completion.Results The head,neck and upper limbs were all fixed desirably by this method,which shortened the operation time,saved 4% paraformaldehyde,and achieved satisfactory immunofluorescence results as compared with conventional procedures.Conclusion The modified method is simple,obviously improves the results of perfusion fixation of rat brain tissue,and therefore deserves to be popularized.

关 键 词：大鼠 脑 灌注固定 免疫荧光 

分 类 号：R743[医药卫生—神经病学与精神病学]