The Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
Department of Applied Physics, Osaka University, Osaka, Japan.
Nat Methods. 2015 Jun;12(6):515-8. doi: 10.1038/nmeth.3362. Epub 2015 Apr 20.
Fluorescence nanoscopy has revolutionized our ability to visualize biological structures not resolvable by conventional microscopy. However, photodamage induced by intense light exposure has limited its use in live specimens. Here we describe Kohinoor, a fast-switching, positively photoswitchable fluorescent protein, and show that it has high photostability over many switching repeats. With Kohinoor, we achieved super-resolution imaging of live HeLa cells using biocompatible, ultralow laser intensity (0.004 J/cm(2)) in reversible saturable optical fluorescence transition (RESOLFT) nanoscopy.
荧光纳米显微镜技术极大地提高了我们对传统显微镜无法分辨的生物结构进行成像的能力。然而，强光照引起的光损伤限制了其在活标本中的应用。在这里，我们描述了 Kohinoor，一种快速切换的、正光可开关的荧光蛋白，并表明它在多次开关重复中具有很高的光稳定性。利用 Kohinoor，我们在活 HeLa 细胞中实现了超分辨率成像，使用的是生物相容性的超低激光强度（0.004 J/cm(2)) 在可逆饱和光学荧光跃迁（RESOLFT）纳米显微镜技术中。
