Platinum porphyrins as phosphorescent label for time-resolved microscopy
Platinum porphyrins as phosphorescent label for time-resolved microscopy
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DOI：
10.1177/002215549704500911
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发表时间：
1997-09-01
影响因子：
3.2
通讯作者：
Tanke, HJ
中科院分区：
文献类型：
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作者：
deHaas, RR;vanGijlswijk, RPM;Tanke, HJ
我们研究了磷光金属卟啉作为时间分辨显微镜的潜在标记物。在对其物理化学性质（量子产率、摩尔吸光系数、衰减时间）进行光谱分析的基础上，挑选出了最佳候选物。接下来，我们合成了抗体和抗生物素蛋白 - 金属卟啉偶联物。确定了这些免疫试剂在量子产率、衰减时间和生物活性保留方面的最佳F/P比值。然后通过原位杂交和免疫细胞化学方法对这些试剂进行评估，用于展示半抗原标记的DNA探针、膜抗原（CD型）和28S rRNA。所有染色样本都显示出明亮的磷光，使用时间分辨显微镜可以选择性地检测到，特别是当在包埋介质中加入葡萄糖/葡萄糖氧化酶以消耗氧气时。讨论了磷光卟啉在强自发荧光材料（组织切片）中时间分辨检测的应用。
We investigated phosphorescent metalloporphyrins as potential labels for time-resolved microscopy. On the basis of spectroscopic analysis of their physicochemical properties (quantum yield, molar absorption coefficient, decay times) the best candidates were selected. Next, we synthesized antibody and avidin metalloporphyrin conjugates. The optimal F/P ratio with respect to quantum yield, decay time, and retention of biological activity of these immunoreagents was determined. The reagents were then evaluated by in situ hybridization and immunocytochemical procedures for demonstration of hapten-labeled DNA probes, membrane antigens (CD type), and 28S rRNA. All stained samples exhibited bright phosphorescence that could be selectively detected using time-resolved microscopy, especially when glucose/glucose oxidase was added to the embedding medium to deplete oxygen. Applications of time-resolved detection of phosphorescent porphyrins in strongly autofluorescent material (histological sections) are discussed.