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首页> 外文期刊>Journal of Neuroscience Methods >Oxygen-enriched photoconversion of fluorescent dyes by means of a closed conversion chamber.
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Oxygen-enriched photoconversion of fluorescent dyes by means of a closed conversion chamber.

机译：通过封闭的转换室对荧光染料进行富氧的光转换。

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摘要

The goal of the present study was an improvement of the widely used photoconversion technique, which still represents the major approach to the ultrastructural analysis of tissue labelled with fluorescent dyes. Since free access of oxygen to the tissue is essential for the dye-dependent photooxidation of diaminobenzidine (DAB), we attempted to facilitate the process using a closed conversion chamber (CCC), which allows photoconversion in an atmosphere of pure oxygen. Fixed rat tissue samples, containing 4Di-10ASP labelled retinal ganglion cells and Cy3 stained cortical perineuronal nets, were choosen to test the applicability and efficiency of the proposed system. The results are compared to corresponding structures photoconverted without pure oxygen. As a result, the employment of the CCC helps saving up to 50% of time required to achieve a comparable degree of photoconversion. Electron microscopical inspection showed no differences between both approaches regarding the distribution of DAB reaction product. However, probably due to the reduced time of irradiation, the ultrastructural integrity of tissue sometimes appeared considerably less affected after photoconversion in the CCC. Additionally, the chamber allowed for safety measures in handling DAB, as the unintentional emission of the presumable carcinogenic substance was completely avoided.
机译：本研究的目的是对广泛使用的光转换技术的改进，该技术仍然代表了用荧光染料标记的组织的超微结构分析的主要方法。由于氧气自由进入组织对于二氨基联苯胺（DAB）的染料依赖性光氧化是必不可少的，因此我们尝试使用封闭的转换室（CCC）简化该过程，该转换室允许在纯氧气氛中进行光转换。选择含有4Di-10ASP标记的视网膜神经节细胞和Cy3染色的皮质神经周围神经网的固定大鼠组织样本，以测试该系统的适用性和效率。将结果与没有纯氧进行光转换的相应结构进行比较。结果，使用CCC有助于节省多达50％的时间，以达到可比的光转换程度。电子显微镜检查显示两种方法在DAB反应产物的分布方面没有差异。但是，可能是由于减少了辐照时间，因此在CCC中进行光转换后，组织的超微结构完整性有时似乎受到的影响较小。另外，由于完全避免了可能的致癌物质的无意排放，该室允许在处理DAB时采取安全措施。

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