Elberger A J, Honig M G
Department of Anatomy and Neurobiology, The University of Tennessee, Memphis 38163.
J Histochem Cytochem. 1990 May;38(5):735-9. doi: 10.1177/38.5.2110209.
The fluorescent carbocyanine dye DiI can be used for retrograde and anterograde labeling of neuronal pathways. To investigate the possible neurochemical identity of DiI-labeled neuronal cell bodies and terminals, we used a procedure for double-labeling of the same tissue with antisera to specific neuroactive substances. This procedure involves visualizing the immunohistochemical label with an FITC-conjugated secondary antiserum. Both labels can be viewed in the same tissue by fluorescence microscopy, and individual cell bodies and processes double-labeled with DiI and antiserum can be identified by switching between filter sets appropriate for rhodamine (to see the DiI labeling) and for fluorescein (to see the immunhistochemical labeling). The method has been used with primary antisera to excitatory and inhibitory amino acid neurotransmitters, as well as to neuropeptides, and is likely to be useful with antibodies against a wide variety of substances. Several other immunocytochemical methods were found to be incompatible with DiI labeling.
荧光碳菁染料DiI可用于神经元通路的逆行和顺行标记。为了研究DiI标记的神经元细胞体和终末可能的神经化学特性，我们采用了一种用针对特定神经活性物质的抗血清对同一组织进行双重标记的方法。该方法包括用异硫氰酸荧光素（FITC）偶联的二抗使免疫组织化学标记可视化。两种标记均可通过荧光显微镜在同一组织中观察到，通过在适合罗丹明（用于观察DiI标记）和异硫氰酸荧光素（用于观察免疫组织化学标记）的滤光片组之间切换，可以识别用DiI和抗血清双重标记的单个细胞体和突起。该方法已用于针对兴奋性和抑制性氨基酸神经递质以及神经肽的一抗，并且可能适用于针对多种物质的抗体。我们发现其他几种免疫细胞化学方法与DiI标记不兼容。
