Rasnik Ivan, McKinney Sean A, Ha Taekjip
Department of Physics, Emory University, Atlanta, Georgia 30322, USA.
Nat Methods. 2006 Nov;3(11):891-3. doi: 10.1038/nmeth934. Epub 2006 Oct 1.
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.
荧光团的光漂白和闪烁对单分子荧光测量的信息含量构成了根本性限制。Cy5的光致闪烁阻碍了许多先前使用这种常用荧光团的研究。在这里，我们表明，Trolox与酶促氧清除系统相结合可消除Cy5闪烁，显著降低光漂白，并在高激发速率下改善信号线性，从而显著扩展了单分子荧光技术的适用性。
