生物重要生物标记物的超分辨率成像的光色光探测器策略
Xianzhi Chai1, Hai-Hao Han1,2, Adam C Sedgwick3
1Key Laboratory for Advanced Materials and Joint International Research Laboratory of Precision Chemistry and Molecular Engineering, Feringa Nobel Prize Scientist Joint Research Center, School of Chemistry and Molecular Engineering, Frontiers Center for Materiobiology and Dynamic Chemistry, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, P. R. China.
Journal of the American Chemical Society
|September 21, 2020
概括
一种新型的β-银酸酶响应探针 (NpG) 与人血清白蛋白 (NpG@HSA) 形成混合体,使细胞成像得到增强. 在β- 银酶激活后,它可以使用STORM成像精确的纳米度可视化酶活性.
科学领域:
- 生物医学工程
- 化学生物学
- 显微镜
背景情况:
- 开发响应的光探测器对于可视化生物过程至关重要.
- 光色探测器具有独特的光学特性,但在水环境和细胞成像中经常面临挑战.
- 人体血清白蛋白 (HSA) 是提高探针溶解性和细胞透性的潜在载体.
研究的目的:
- 设计和表征一个对β-galactosidase (β-Gal) 敏感的光色光探针 (NpG),该探针可以预先结合HSA.
- 开发一种新型的成像平台 (NpG@HSA),用于增强β-Gal活动的细胞吸收和可视化.
- 使用STORM显微镜对β-Gal活性进行高分辨率亚细胞定位.
主要方法:
- 一个对β-galactosidase有反应的光色光探针 (NpG) 的设计,其中包含一个纳夫他胺和一个螺旋单元.
- 通过预先将NpG与人血清白蛋白结合,形成探针/蛋白混合物 (NpG@HSA).
- 使用光显微镜和STORM可视化探针吸收和β-Gal活动的细胞成像实验.
- 用于先进的成像能力的梅洛胺形式的光异构化研究.
主要成果:
- NpG@HSA混合体表现出增强的光发射 (520 nm) 和改善的细胞透性和可溶性.
- 通过β-Gal介导的NpG@HSA分裂导致红色光转移 (620 nm) 和NpM@HSA的形成.
- 使用NpG@HSA进行的STORM成像在确定β-Gal活动的亚细胞分布方面实现了纳米级精度.
- 该探测器表现出极少的光毒性,优异的光稳定性和可逆性的高级显微镜.
结论:
- NpG@HSA系统为光色光探测器提供了多功能成像平台.
- 这种方法可以精确可视化酶活性和生物标志物的亚细胞定位.
- 这项研究突显了设计用于细胞过程中特定疾病生物标志物的先进探测器的潜力.
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