基于可光切换的拉曼探测器的超高分辨率振动成像
Jingwen Shou1, Ayumi Komazawa2, Yuusaku Wachi3
1Department of Electrical Engineering and Information Systems, The University of Tokyo, Tokyo 113-8656, Japan.
Science advances
|June 16, 2023
概括
可逆和光学拉曼转换 (RESORT) 显微镜可实现活细胞的超分辨率振动成像. 这种新的技术克服了当前方法的局限性,提供高化学特异性和超出光学衍射极限的空间分辨率.
科学领域:
- 生物光子学 生物光子学
- 化学成像技术 化学成像技术
- 细胞显微镜 细胞显微镜
背景情况:
- 超分辨率的振动显微镜提供了多重纳米尺度生物成像的潜力,因为与光相比,分子振动的光谱线宽较窄.
- 现有的超高分辨率振动显微镜技术面临诸如需要细胞固定,高激光功率要求和复杂的检测设置等局限性.
研究的目的:
- 引入可逆和光学拉曼转换 (RESORT) 显微镜,这是一种旨在克服当前超分辨率振动显微镜局限性的新技术.
- 为了证明 RESORT 显微镜的有效性,使用可光切换的拉曼探针对活细胞进行高分辨率成像.
主要方法:
- 开发和验证使用低功率连续波激光光的可控制信号激活和消耗的明亮光交换拉曼探测器 (DAE620).
- 通过利用可光切换刺激拉曼散射 (SRS) 通过甜甜圈形光束对DAE620的信号耗尽来实现超高分辨率成像.
主要成果:
- 用RESORT显微镜在哺乳动物细胞中进行超分辨率振动成像的演示.
- 取得了卓越的化学特异性和空间分辨率,超过了光学衍射极限.
- 在低功率激光照明下验证了DAE620探针的可光切换特性.
结论:
- RESORT显微镜有效地克服了现有的超高分辨率振动成像技术的局限性.
- 开发的可光切换探头和RESORT方法显示,对活生物样本的多重超分辨率成像具有很高的潜力.
- 这种技术为先进的细胞成像提供了一个有前途的新工具,具有高化学特异性和分辨率.
相关概念视频
01:26Raman Spectroscopy Instrumentation: Overview
468
A conventional Raman spectrophotometer includes a laser source, a sample holding system, a wavelength selector, and a detector.
The monochromatic laser source, typically using visible or near-infrared radiation, generates a highly focused beam of light. This light interacts with the molecules of the sample, scattering some of the light. Liquid and gaseous samples are usually tested in ordinary glass capillaries, while solids can be analyzed as powders packed in capillaries or as potassium...
The monochromatic laser source, typically using visible or near-infrared radiation, generates a highly focused beam of light. This light interacts with the molecules of the sample, scattering some of the light. Liquid and gaseous samples are usually tested in ordinary glass capillaries, while solids can be analyzed as powders packed in capillaries or as potassium...
468
01:20Raman Spectroscopy: Overview
474
The underlying principle of Raman spectroscopy is based on the interaction between light and matter, specifically molecules' inelastic scattering of photons. When a monochromatic beam of light, typically from a laser source, interacts with a sample, most scattered light has the same frequency as the incident light. This is known as Rayleigh scattering.
However, a small fraction of the scattered light exhibits a frequency shift due to the exchange of energy between the incident photons and...
However, a small fraction of the scattered light exhibits a frequency shift due to the exchange of energy between the incident photons and...
474
01:37Super-resolution Fluorescence Microscopy
7.1K
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...
7.1K