Fredman S M
J Neurosci Methods. 1987 Jul;20(3):181-94. doi: 10.1016/0165-0270(87)90050-1.
A new method of intracellularly staining neurons is described. Nickel-lysine (NL) can be used for both intracellular injection by pressure or iontophoresis and retrograde labelling (axonal backfilling). Once introduced into neurons, NL is reacted with dithiooximide dissolved in dimethyl sulfoxide (DMSO) to produce a blue-black precipitate. Small diameter processes are easily detected. For pressure injections, mixing NL with carboxyfluorescein provides a simple way to gauge how much dye has been injected, in that the latter is readily visible when illuminated with blue light. NL appears to move within neurons by axonal transport. Staining over long distances can be obtained in 12-24 h. NL does not appear to cross electrotonic synapses and remains confined to the neurons into which it has been injected. NL staining is simple, flexible and inexpensive. It has the additional advantage that it is compatible with other staining techniques.
本文描述了一种新的细胞内染色神经元的方法。镍赖氨酸（NL）可用于通过压力或离子电泳进行细胞内注射以及逆行标记（轴突回充）。一旦将NL引入神经元，它会与溶解在二甲基亚砜（DMSO）中的二硫代肟反应，产生蓝黑色沉淀。小直径的突起很容易被检测到。对于压力注射，将NL与羧基荧光素混合提供了一种简单的方法来衡量注射了多少染料，因为在用蓝光照射时，后者很容易被看到。NL似乎通过轴突运输在神经元内移动。在12 - 24小时内可以获得长距离的染色。NL似乎不会穿过电突触，并且仍局限于注射了它的神经元内。NL染色简单、灵活且成本低廉。它还有一个额外的优点，即它与其他染色技术兼容。
