细胞代谢的无标签纳米显微镜通过超敏感重重可见刺激拉曼散射
Haonan Lin1,2,3, Scott Seitz4,5, Yuying Tan2,3
1Department of Electrical and Computer Engineering, Boston University, Boston, MA, USA.
Nature methods
|January 17, 2025
概括
我们开发了超敏感重权可见刺激拉曼散射 (URV-SRS) 显微镜,用于无标签的细胞代谢成像. 该技术实现了纳米分辨率,使细胞和细菌中代谢纳米结构的可视化成为可能.
科学领域:
- 化学成像技术 化学成像技术
- 细胞的新陈代谢
- 纳米显微镜使用的方法.
背景情况:
- 目前的超分辨率成像方法在可视化小代谢物方面面临局限性,原因是染料不兼容和分辨率有限.
- 图像质谱仪为详细的代谢分析提供有限的空间分辨率.
研究的目的:
- 开发一种新的无标签振动成像技术,用于细胞内代谢物的多重纳米镜.
- 克服现有的细胞代谢高分辨率成像方法的局限性.
主要方法:
- 开发了一种可见刺激拉曼散射 (SRS) 显微镜,具有广泛的脉冲声.
- 实现自主监督的多剂消噪器,以抑制SRS信号中的噪声.
- 应用富里埃重量调整来增强100nm以下的空间频率.
主要成果:
- 达到大约4000个分子的检测极限,比近红外SRS提高了50倍的灵敏度.
- 使用开发的消噪器,抑制非独立噪声超过7.2dB.
- 在细胞成像中达到86nm的横向分辨率,通过富里埃环相关性验证.
结论:
- 超敏感重权可见刺激拉曼散射 (URV-SRS) 能够实现纳米空间代谢.
- 在工程细菌中证明了病毒复制和脂肪酸合成期间代谢纳米结构的可视化.
- URV-SRS提供了一种强大的无标签方法,用于高分辨率的细胞代谢成像.
相关概念视频
01:37Super-resolution Fluorescence Microscopy
Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.
01:25Overview of Electron Microscopy
The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.