我们发现最常引起急性分离神经元失败的主要原因有实验用水污染、溶液浓度错误、处理组织块时手法粗暴。当分离新皮层以外的神经元或大鼠以外的其他哺乳动物神经元时，应反复试验，以明确最佳的消化时间。在分离某些脑区神经元时，蛋白酶K并不是必需的。
对组织进行分离吹打时没有什么特殊的技巧，只是需缓慢、轻柔的将组织吸入细管，随后缓缓的吹出，在此过程中避免气泡的产生。吹打时没有必要完全打碎所有的组织。组织在经过大口的巴氏细管几次后就可以释放出神经元。研究者在试验时应根据自己所用的组织凭经验确定出最适的吹打方法。
预期结果
100-150g体重范围内的大鼠，每毫克新皮层能够产生730 225 (mean SD, n = 7)个神经元；
275-325g体重范围内的大鼠，每毫克新皮层能够产生460 (mean SD, n = 6)个神经元。
实验时间
从动物断头取脑到分离出第一批神经元需要45分种。神经元分离出后应立即着手进行相关电生理操作。
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供稿：Alan R. Kay和David J. Krupa 爱荷华大学，爱荷华城，爱荷华州，美国
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