FURTHER-STUDIES ON THE USE OF THE FLUORESCENT TRACERS FAST BLUE AND DIAMIDINO YELLOW - EFFECTIVE UPTAKE AREA AND CELLULAR-STORAGE SITES
FURTHER-STUDIES ON THE USE OF THE FLUORESCENT TRACERS FAST BLUE AND DIAMIDINO YELLOW - EFFECTIVE UPTAKE AREA AND CELLULAR-STORAGE SITES
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DOI：
10.1016/0165-0270(87)90100-2
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发表时间：
1987-09-01
影响因子：
3
通讯作者：
CONDE, F
中科院分区：
文献类型：
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作者：
CONDE, F
利用猫小脑红色核向前间核（NIA）的投影，研究了荧光示踪剂Fast blue （FB）和Diamidino yellow （DY）使用的一些基本方法学问题。在标准的nissl染色切片上，可以在FB和DY注射部位划定4个不同的区域。对NIA注射部位和对侧红核细胞体逆行标记的相关研究表明，有效摄取仅发生在被注射针机械损伤的区域（称为0区）。在注射后存活期间，示踪剂保持在该区域。两种示踪剂的有限摄取区域是研究限制性神经元投射的一个有价值的特征。然而，这些示踪剂不适合用于定量研究，特别是那些涉及轴突侧支的研究。用水溶性固定剂灌注不改变细胞储存部位。在使用辣根过氧化物酶和DY的双重标记实验中，HRP组织化学诱导了DY的重要“冲洗”，因此，标记神经元的数量被低估。
Some basic methodological issues concerning the use of the fluorescent tracers Fast blue (FB) and Diamidino yellow (DY) were studied using the projections of the red nucleus to the nucleus interpositus anterior (NIA) of the cerebellum of the cat. On standard Nissl-stained sections, it was possible to delineate 4 distinct zones at the FB and DY injection sites. Correlative studies of injection sites in the NIA and retrograde labeling of cell bodies in the contralateral red nucleus showed that effective uptake occurred only from the zone mechanically damaged by the injection needle (termed zone 0). The tracer remains in this zone during the post-injection survival. The limited uptake area for both tracers is a valuable feature for studies of restricted neuronal projections. However, the tracers are not suitable for use in quantitative studies, especially those concerning axonal collateralization. Perfusion with water-soluble fixatives did not alter the cellular storage site. In double-labeling experiments using horseradish peroxidase and DY, the HRP histochemistry induced an important "washing out" of DY and consequently, an underestimation of the number of labeled neurons.