CRISPR-PAINT nanorulers

The GATTA-PAINT 40 RG nanoruler (GATTAquant, DNA nanotechnologies) consists of three fluorophores (ATTO 655/ATTO 542) separated 40 nm from each other. Imaging of the sample was performed on an Olympus IX-81 inverted microscope equipped with an Olympus UApo N 100X/1.49 NA, oil-immersion objective in TIRF illumination mode (Olympus, cellTIRF Illuminator) with ~200 nm evanescence field depth. Sample excitation was provided by 488 nm and 561 nm lasers at 23.1 mW laser power. 300 total images were collected with an EM-CCD sensor (Andor iXon 897), exposure time set to 50 ms per frame and a pixel size of 160 nm.

GATTA-PAINT 40 RG纳米标尺（GATTAquant，DNA纳米技术领域）由三个间距为40 nm的荧光基团（ATTO 655/ATTO 542）构成。该样本的成像实验在奥林巴斯IX-81倒置显微镜上完成，该显微镜配备奥林巴斯UApo N 100X/1.49 NA油浸物镜，采用全内反射荧光（Total Internal Reflection Fluorescence, TIRF）照明模式（搭载奥林巴斯cellTIRF Illuminator照明器），消逝场深度约为200 nm。为样本提供激发的是功率为23.1 mW的488 nm与561 nm激光。实验共采集300幅图像，使用Andor iXon 897型电子倍增电荷耦合器件（Electron Multiplying Charge-Coupled Device, EM-CCD）传感器，单帧曝光时长设置为50 ms，像素尺寸为160 nm。