Clancy B, Cauller L J
Department of Psychology, Cornell University, Ithaca, NY 14853, USA.
J Neurosci Methods. 1998 Sep 1;83(2):97-102. doi: 10.1016/s0165-0270(98)00066-1.
Autofluorescence of aldehyde-fixed neural tissue often obscures perikaria and fine processes labeled with fluorescent anterograde or retrograde tracers. In particular, this autofluorescence hinders the detectability of fine axonal projections labeled with the convenient anterograde tracer, tetramethylrhodamine dextranamine. Background fluorescence was notably reduced by immersion of free-floating brain tissue sections in a solution of sodium borohydride (NaBH4, 0.1%), a chemical which is known to neutralize Schiffs bases through reduction of amine-aldehyde compounds into non-fluorescent salts. The reversible and renewable immersion technique was most effective in paraformaldehyde-fixed tissue where the preservation quality was improved such that labeled axons remained detectable for more than 1 year after initial preparation.
醛固定神经组织的自发荧光常常会掩盖被荧光顺行或逆行示踪剂标记的胞体和细微突起。特别是这种自发荧光会妨碍使用便捷的顺行示踪剂四甲基罗丹明葡聚糖胺标记的细微轴突投射的可检测性。通过将游离的脑组织切片浸入硼氢化钠(NaBH4,0.1%)溶液中,背景荧光显著降低,硼氢化钠是一种已知可通过将胺醛化合物还原为非荧光盐来中和席夫碱的化学物质。这种可逆且可再生的浸泡技术在多聚甲醛固定的组织中最为有效,在这种组织中保存质量得到改善,以至于在初始制备后1年多仍可检测到标记的轴突。