Super-beacons: Open-source probes with spontaneous tuneable blinking compatible with live-cell super-resolution microscopy.,Traffic

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Super-beacons: Open-source probes with spontaneous tuneable blinking compatible with live-cell super-resolution microscopy.
Traffic ( IF 3.6 ) Pub Date : 2020-03-30 , DOI: 10.1111/tra.12728
Pedro M Pereira _{1,

2,

3} , Nils Gustafsson ₁ , Mark Marsh ₁ , Musa M Mhlanga ₄ , Ricardo Henriques _{1,

2}

Affiliation

Localization-based super-resolution microscopy relies on the detection of individual molecules cycling between fluorescent and non-fluorescent states. These transitions are commonly regulated by high-intensity illumination, imposing constrains to imaging hardware and producing sample photodamage. Here, we propose single-molecule self-quenching as a mechanism to generate spontaneous photoswitching. To demonstrate this principle, we developed a new class of DNA-based open-source super-resolution probes named super-beacons, with photoswitching kinetics that can be tuned structurally, thermally and chemically. The potential of these probes for live-cell compatible super-resolution microscopy without high-illumination or toxic imaging buffers is revealed by imaging interferon inducible transmembrane proteins (IFITMs) at sub-100 nm resolutions.

中文翻译：

超级信标：具有自发可调闪烁的开源探针，与活细胞超分辨率显微镜兼容。

基于定位的超分辨率显微镜依赖于检测荧光状态和非荧光状态之间循环的单个分子。这些转变通常由高强度照明调节，对成像硬件施加限制并产生样本光损伤。在这里，我们提出单分子自猝灭作为产生自发光开关的机制。为了证明这一原理，我们开发了一种新型基于 DNA 的开源超分辨率探针，称为超级信标，其光开关动力学可以在结构、热和化学方面进行调整。通过以亚 100 nm 分辨率对干扰素诱导跨膜蛋白 (IFITM) 进行成像，揭示了这些探针在无需高照度或有毒成像缓冲液的情况下用于活细胞兼容超分辨率显微镜的潜力。

更新日期：2020-04-22

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