小鼠脑切片漂浮染色方法研究

Research on free-floating staining of mouse brain slides

为了提高小鼠脑组织学研究效率,解决传统石蜡切片和冰冻切片样品准备时间长、步骤复杂等问题,提出了振动切片结合漂浮染色快速进行小鼠脑组织切片形态学研究的方法。该方法通过小鼠心脏灌注后,获取脑组织,利用琼脂糖快速包埋固定;通过振动切片方法,40微米厚度连续获取脑组织切片,结合漂浮染色观测特征区域神经元形态;最后通过体视学方法,快速统计估算神经元数量。实验结果表明,振动切片联合漂浮染色可快速进行小鼠脑组织学研究,并具有较高的稳定性和可重复性。

To improve the efficiency of brain histological research in mouse,this study deigned a new method that using free-floating staining post vibratome section.The new method can avoid the long-timing preparation and complicated process in paraffin and frozen section.In this method,we use agarose to assemble the brain after the post-cardiac perfusion.Forty μm brain slides were continuously collected by vibratome section,and the slides were developed with anti-NeuN by free-floating staining,then the neuron number could be rapidly calculated by stereological method.Our method indicated a stable and repeatable brain section and staining protocol that using both vibratome section and free-floating staining.