通过双色超分辨率光学涨落成像（SOFI）解析细胞内组分之间的空间关系。
Resolving the spatial relationship between intracellular components by dual color super resolution optical fluctuations imaging (SOFI).
作者信息
Gallina Maria Elena, Xu Jianmin, Dertinger Thomas, Aizer Adva, Shav-Tal Yaron, Weiss Shimon
机构信息
Department of Chemistry and Biochemistry, Department of Physiology, and the California NanoSystem Institute (CNSI), University of California Los Angeles, Los Angeles, CA 90095-1569, USA ; Department of Chemistry "G. Ciamician", University of Bologna, Via Selmi 2, Bologna 40126, Italy.
出版信息
Opt Nanoscopy. 2013 Feb 25;2. doi: 10.1186/2192-2853-2-2.
BACKGROUND
Multi-color super-resolution (SR) imaging microscopy techniques can resolve ultrastructura relationships between- and provide co-localization information of- different proteins inside the cell or even within organelles at a higher resolution than afforded by conventional diffraction-limited imaging. While still very challenging, important SR colocalization results have been reported in recent years using STED, PALM and STORM techniques.
RESULTS
In this work, we demonstrate dual-color Super Resolution Optical Fluctuations Imaging (SOFI) using a standard far-field fluorescence microscope and different color blinking quantum dots. We define the spatial relationship between hDcp1a, a processing body (P-body, PB) protein, and the tubulin cytoskeletal network. Our finding could open up new perspectives on the role of the cytoskeleton in PB formation and assembly. Further insights into PB internal organization are also reported and discussed.
CONCLUSIONS
Our results demonstrate the suitability and facile use of multi-color SOFI for the investigation of intracellular ultrastructures.
背景
多色超分辨率（SR）成像显微镜技术能够解析细胞内甚至细胞器内不同蛋白质之间的超微结构关系，并提供共定位信息，其分辨率高于传统衍射极限成像。尽管仍然极具挑战性，但近年来利用受激发射损耗（STED）、光激活定位显微镜（PALM）和随机光学重建显微镜（STORM）技术已报道了重要的SR共定位结果。
结果
在本研究中，我们使用标准远场荧光显微镜和不同颜色闪烁量子点展示了双色超分辨率光学涨落成像（SOFI）。我们确定了处理小体（P小体，PB）蛋白hDcp1a与微管蛋白细胞骨架网络之间的空间关系。我们的发现可能为细胞骨架在PB形成和组装中的作用开辟新的视角。还报道并讨论了对PB内部组织的进一步见解。
结论
我们的结果证明了多色SOFI在研究细胞内超微结构方面的适用性和易用性。
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