Nonblinking and longlasting single-molecule fluorescence imaging
Nonblinking and longlasting single-molecule fluorescence imaging
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DOI：
10.1038/nmeth934
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发表时间：
2006-11-01
期刊：
影响因子：
48
通讯作者：
Ha, Taekjip
中科院分区：
文献类型：
--
作者：
Rasnik, Ivan;McKinney, Sean A.;Ha, Taekjip
荧光团的光漂白和闪烁对单分子荧光测量的信息量构成了根本的限制。Cy5的光诱导闪烁阻碍了许多以前使用这种流行的荧光团进行的研究。在这里，我们表明，Trolox与酶促氧清除系统相结合，消除了Cy5闪烁，显著减少了光漂白，并在高激发速率下改善了信号线性度，显著扩展了单分子荧光技术的适用范围。
Photobleaching and blinking of fluorophores pose fundamental limitations on the information content of single-molecule fluorescence measurements. Photoinduced blinking of Cy5 has hampered many previous investigations using this popular fluorophore. Here we show that Trolox in combination with the enzymatic oxygen-scavenging system eliminates Cy5 blinking, dramatically reduces photobleaching and improves the signal linearity at high excitation rates, significantly extending the applicability of single-molecule fluorescence techniques.