resPAINT:通过积极控制探测器发射来加速体积超分辨率定位显微镜
Edward W Sanders1, Alexander R Carr1, Ezra Bruggeman1
1Yusuf Hamied Department of Chemistry University of Cambridge Cambridge CB2 1EW UK.
Angewandte Chemie (Weinheim an der Bergstrasse, Germany)
|March 20, 2024
概括
储-PAINT (resPAINT) 通过积累探头来增强超高分辨率显微镜,实现50倍更高的定位率. 这一突破使得整个细胞表面和膜蛋白的详细成像成为可能.
科学领域:
- 生物物理学的生物物理.
- 显微镜的使用方法
- 纳米技术 纳米技术
背景情况:
- 像PAINT这样的定位显微镜技术提供了高精度,但面临着局限性.
- 在PAINT中探针度高导致背景光,阻碍体积成像.
- 现有的方法在不损害图像对比度的情况下努力实现高本地化率.
研究的目的:
- 开发一种先进的PAINT技术,改进探头积累和光物理控制.
- 为了克服传统PAINT的局限性,特别是在高探头度场景中.
- 为了使生物结构的高分辨率成像,包括整个细胞表面和膜蛋白.
主要方法:
- 引入了储PAINT (resPAINT),将PAINT与探测器光物理学的主动控制相结合.
- 开发了一种可激活的探测器"储",它会积聚在目标分子上.
- 结合resPAINT与大深度显微镜进行体积超分辨率成像.
主要成果:
- 与传统PAINT相比,本地化率提高了50倍.
- 保持高图像对比度,尽管增加了探头积累.
- 使用resPAINT.显示了整个细胞表面的超高分辨率成像.
- 通过使用resPAINT与Fab片段成功成像了膜蛋白.
结论:
- resPAINT显著提高了局部化显微镜的性能.
- 该技术扩大了PAINT用于成像各种生物相互作用和结构的适用性.
- resPAINT为生物研究中的高分辨率,高速成像提供了一个强大的工具.
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