Two rapid methods of counterstaining fluorescent dye tracer containing sections without reducing the fluorescence.
Two rapid methods of counterstaining fluorescent dye tracer containing sections without reducing the fluorescence.
复制标题
两种快速复染含有荧光染料示踪剂的切片而不降低荧光的方法。
DOI：
10.1016/0006-8993(86)91375-2
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发表时间：
1986
期刊：
影响因子：
2.9
通讯作者：
Wyss,JM
中科院分区：
文献类型：
--
作者：
Sripanidkulchai,K;Wyss,JM
描述了一种含有被荧光染料或辣根过氧化物酶(HRP)逆行标记的细胞的神经组织的反染色方法。具体地说，详细说明了示踪剂与亚甲基蓝联合用于尼氏染色或与银法联合使用以检测乙酰胆碱酯酶的方案。根据皮质-皮质和丘脑-皮质投影来评估这些技术的有效性。结果表明，这些方法不会掩盖最敏感的荧光染料的标记或HRP的标记。只有黄色染料会受到亚甲基蓝反染的显著影响。此外，神经元中的快速蓝标记并不会因Bodian纤维染色而显著减少。还评估了包含荧光染料标记细胞的盖片切片的效果，发现在不降低灵敏度的情况下显著延长了标记的寿命。因此，这两种反染色技术提供了很好的结构信息，不会严重影响示踪剂标记，并且几乎没有其他反染色方法所共有的缺点。
A method is described for counterstaining neural tissue containing cells that are retrogradely labeled by flourescent dyes or horseradish peroxidase (HRP). Specifically, protocols are detailed for the combined use of the tracers with Methylene blue for a Nissl stain or with silver methods for the detection of acetylcholine esterase. The usefulness of these techniques is evaluated in relation to cortico-cortico and thalamocortico projections. The findings indicate that the methods do not mask the labeling of the most sensitive fluorescent dyes or by HRP. Only the yellow dyes are significantly affected by the Methylene blue counterstain. Further, Fast blue labeling in neurons is not significantly diminished by the Bodian fiber stain. The effect of coverslipping sections containing fluorescent dye labeled cells also was evaluated and found to significantly extend the life of the labeling while not reducing the sensitivity. Thus the two counterstaining techniques provide excellent structural information, do not seriously affect tracer labeling and have few of the disadvantages common to other counterstaining methods.