Zhang Dong, Liu Shi Gang, Fu Zhaodi, He Yu, Gao Wenli, Shi Xingbo
Laboratory of Micro and Nano Biosensing Technology in Food Safety, Hunan Provincial Key Laboratory of Food Science and Biotechnology, College of Food Science and Technology, Hunan Agricultural University, Changsha 410128, China.
Analytical Testing Laboratory, Changsha Research Institute of Mining and Metallurgy CO., LTD., Changsha 410012, China.
MethodsX. 2020 Nov 20;7:101155. doi: 10.1016/j.mex.2020.101155. eCollection 2020.
• Smart usage of single molecule photobleaching technology and dual-color fluorescence colocalization is of critical importance for exploiting the sensing platform. Here, we provide the detailed protocols related to the article "A split aptamer sensing platform for highly sensitive detection of theophylline based on dual-color fluorescence colocalization and single molecule photobleaching" (published online by Biosensors and Bioelectronics) (Liu et al., 2020). The protocols contain: (1) how to clean the slides; (2) how to prepare the probe and detection sample; (3) Single molecule imaging; 4) Data processing by using the Image J. Finally, we used a simple model to confirm the feasibility of the method for integrating dual-color fluorescence colocalization and single molecule photobleaching technology on the theophylline sensing platform. • A simple, ultrasensitive method for the detection of theophylline. • The method is easily comprehensible. • Both strategy formulation and data processing are simple, learnability, and highly reproducible.
• 巧妙运用单分子光漂白技术和双色荧光共定位对于开发传感平台至关重要。在此，我们提供了与文章《基于双色荧光共定位和单分子光漂白的用于高灵敏检测茶碱的分裂适体传感平台》（发表于《生物传感器与生物电子学》在线版）（Liu等人，2020年）相关的详细实验方案。这些方案包括：（1）如何清洁载玻片；（2）如何制备探针和检测样品；（3）单分子成像；（4）使用Image J进行数据处理。最后，我们使用一个简单模型证实了在茶碱传感平台上整合双色荧光共定位和单分子光漂白技术的方法的可行性。
• 一种用于检测茶碱的简单、超灵敏方法。
• 该方法易于理解。
• 策略制定和数据处理都简单、易学且具有高度可重复性。
