Ratiometric Sensing of Redox Environments Inside Individual Carboxysomes Trapped in Solution.
Ratiometric Sensing of Redox Environments Inside Individual Carboxysomes Trapped in Solution.
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DOI：
10.1021/acs.jpclett.2c00782
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发表时间：
2022-05-26
影响因子：
5.7
通讯作者：
Moerner, W. E.
中科院分区：
文献类型：
--
作者：
Carpenter, William B.;Lavania, Abhijit A.;Borden, Julia S.;Oltrogge, Luke M.;Perez, Davis;Dahlberg, Peter D.;Savage, David F.;Moerner, W. E.
生物纳米颗粒在溶液中的扩散，阻碍了我们对单个颗粒进行连续监测并测量其物理和化学性质的能力。为解决这一问题，我们此前研发了干涉散射反布朗电动（ISABEL）光镊，它利用散射来定位颗粒，并施加与布朗运动相抗衡的电动作用力，从而实现长时间观测。在此，我们介绍一种改进的ISABEL光镊，它整合了近红外散射照明光束，并能快速交替使用405纳米和488纳米的荧光激发报告光束。借助ISABEL光镊，我们监测了用比率型氧化还原报告分子roGFP2标记的单个羧酶体内部的氧化还原环境。羧酶体在散射对比度（反映尺寸）和依赖氧化还原的比率型荧光方面差异很大。此外，我们利用氧化还原传感技术探究了完整羧酶体内的化学动力学，而整体测量可能会受到聚集体或干扰性荧光蛋白的不良影响。总体而言，我们展示了ISABEL光镊灵敏监测纳米级生物物体的能力，为这些系统开展新实验创造了条件。
Diffusion of biological nanoparticles in solution impedes our ability to continuously monitor individual particles and measure their physical and chemical properties. To overcome this, we previously developed the interferometric scattering anti-Brownian electrokinetic (ISABEL) trap, which uses scattering to localize a particle and applies electrokinetic forces that counteract Brownian motion, thus enabling extended observation. Here we present an improved ISABEL trap that incorporates a near-infrared scatter illumination beam and rapidly interleaves 405 and 488 nm fluorescence excitation reporter beams. With the ISABEL trap, we monitored the internal redox environment of individual carboxysomes labeled with the ratiometric redox reporter roGFP2. Carboxysomes widely vary in scattering contrast (reporting on size) and redox-dependent ratiometric fluorescence. Furthermore, we used redox sensing to explore the chemical kinetics within intact carboxysomes, where bulk measurements may contain unwanted contributions from aggregates or interfering fluorescent proteins. Overall, we demonstrate the ISABEL trap’s ability to sensitively monitor nanoscale biological objects, enabling new experiments on these systems.
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DOI：
10.1073/pnas.0601210103
发表时间：
2006-05-30
影响因子：
11.1
作者：
Friedel, Miriam;Baumketner, Andrij;Shea, Joan-Emma
通讯作者：
Shea, Joan-Emma
影响因子：
1.2
作者：
Gemeinhardt, Andre;McDonald, Matthew P.;Sandoghdar, Vahid
通讯作者：
Sandoghdar, Vahid
影响因子：
21.8
作者：
Goldsmith, Randall H.;Moerner, W. E.
通讯作者：
Moerner, W. E.
影响因子：
2.9
作者：
Oltrogge LM;Wang Q;Boxer SG
通讯作者：
Boxer SG
影响因子：
4.8
作者：
Dou, Zhicheng;Heinhorst, Sabine;Cannon, Gordon C.
通讯作者：
Cannon, Gordon C.