Dynamic three-dimensional tracking of single fluorescent nanoparticles deep inside living tissue.
Dynamic three-dimensional tracking of single fluorescent nanoparticles deep inside living tissue.
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DOI：
10.1364/oe.20.019697
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发表时间：
2012-08
期刊：
影响因子：
3.8
通讯作者：
Jan-Hendrik Spille;Tim P. Kaminski;Heinz-Peter Königshoven;U. Kubitscheck
中科院分区：
文献类型：
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作者：
Jan-Hendrik Spille;Tim P. Kaminski;Heinz-Peter Königshoven;U. Kubitscheck
通过像散成像可以将三维(3D)空间信息编码到荧光纳米颗粒的二维图像中。我们将这种方法与光片显微镜相结合，用于在活体组织中进行深达200微米的高对比度单粒子成像，并通过实时图像分析来确定具有纳米精度和毫秒时间分辨率的3D粒子定位。轴向信息被立即引导到样本台，以保持焦平面内运动的粒子处于主动反馈环路中。我们展示了对纳米粒子的3D跟踪，在数千帧的大细胞核中以前所未有的深度进行跟踪，每个空间维度的范围超过10微米，同时获得光学切片的广域图像。我们的结论是，这种使用光片显微镜的3D粒子跟踪技术是对纳米显微镜工具箱的有价值的扩展。
Three-dimensional (3D) spatial information can be encoded in two-dimensional images of fluorescent nanoparticles by astigmatic imaging. We combined this method with light sheet microscopy for high contrast single particle imaging up to 200 µm deep within living tissue and real-time image analysis to determine 3D particle localizations with nanometer precision and millisecond temporal resolution. Axial information was instantly directed to the sample stage to keep a moving particle within the focal plane in an active feedback loop. We demonstrated 3D tracking of nanoparticles at an unprecedented depth throughout large cell nuclei over several thousand frames and a range of more than 10 µm in each spatial dimension, while simultaneously acquiring optically sectioned wide field images. We conclude that this 3D particle tracking technique employing light sheet microscopy presents a valuable extension to the nanoscopy toolbox.