Pinault D
Université Laval, Centre de Recherches en Neurobiologie, Hôpital de l'Enfant-Jésus, Québec, Canada.
Neurosci Lett. 1994 Apr 11;170(2):255-60. doi: 10.1016/0304-3940(94)90332-8.
We describe a novel and powerful extracellular method for the staining of a single neuron identified by electrophysiological criteria. This single-unit technique involves the use of glass micro-electrodes (tip diameter: 1.5-2.5 microns) filled with a saline solution (NaCl; 0.5 M) containing 1.5% of biocytin or Neurobiotin. Once a neuron is recorded, isolated and identified, the tracer is delivered by anodal current pulses of a few nA. The cell must remain well isolated and alive during the ejection procedure to ensure optimal staining. Evidence is provided that the labeled neuron is actually the one that was recorded. Our simple method is reliable with a success rate exceeding 85%. The advantages and pitfalls are discussed. This single-unit labeling technique could further be combined with any other procedures ranging from biological to behavioral studies.
我们描述了一种新颖且强大的细胞外方法,用于对通过电生理标准鉴定的单个神经元进行染色。这种单细胞技术涉及使用填充有含1.5%生物胞素或神经生物素的盐溶液(氯化钠;0.5M)的玻璃微电极(尖端直径:1.5 - 2.5微米)。一旦记录、分离并鉴定出一个神经元,通过几纳安的阳极电流脉冲来输送示踪剂。在注射过程中,细胞必须保持良好的分离状态且存活,以确保最佳染色效果。有证据表明被标记的神经元实际上就是所记录的那个神经元。我们的简单方法可靠,成功率超过85%。文中讨论了其优点和缺陷。这种单细胞标记技术可进一步与从生物学研究到行为学研究的任何其他程序相结合。