Mieskes Frank, Wehnekamp Fabian, Plucińska Gabriela, Thong Rachel, Misgeld Thomas, Lamb Don C
Department of Chemistry, Center for Nano Science (CENS), Center for Integrated Protein Science (CIPSM), Nanosystems Initiative Muünchen (NIM), Ludwig Maximilians-Universität München, Munich, Germany.
Munich Cluster for Systems Neurology (SNergy), Center for Integrated Protein Science (CIPSM), German Center for Neurodegenerative Diseases (DZNE), Institute of Neuronal Cell Biology, Technische Universitätt München, Munich, Germany.
Data Brief. 2020 Feb 13;29:105280. doi: 10.1016/j.dib.2020.105280. eCollection 2020 Apr.
Recently, a large number of single particle tracking (SPT) approaches have been developed. Generally, SPT techniques can be split into two groups: approaches where trajectory extraction is carried out after data acquisition and approaches that perform particle tracking in real time [1]. One feedback approach is 3D Orbital Tracking, where the laser excitation beam is rotated in a circle about the object, generating a so called orbit [2,3]. By calculating the particle position from the detected intensity after every orbit in relation to its center, this method allows the microscope to follow a single object in real time. The high spatiotemporal resolution of this method and the potential to optically manipulate the followed object during the measurement promises to yield new deep insights into biological systems [4-7]. By upgrading this approach in a way that the specimen is recentered by a xy-stage on the center of the microscope, particle tracking with this long-range tracking feature is no longer limited to the covered field-of-view. This allows for the observation of mitochondrial trafficking in living zebrafish embryos over long distances. Here, we provide the raw data for antero- and retrograde movement of mitochondria labelled with photo-activatable green fluorescent protein (mitoPAGFP). It relates to the scientific article "Nanoresolution real-time 3D orbital tracking for studying mitochondrial trafficking in vertebrate axons in vivo" [8]. By applying a correlation analysis on the trajectories, it is possible to distinguish between active transport and pausing events with less biasing compared to the mean squared displacement approach.
最近,已经开发出了大量的单粒子追踪(SPT)方法。一般来说,SPT技术可以分为两组:在数据采集后进行轨迹提取的方法和实时进行粒子追踪的方法[1]。一种反馈方法是3D轨道追踪,其中激光激发光束围绕物体做圆周旋转,产生所谓的轨道[2,3]。通过在每个轨道之后根据检测到的强度相对于其中心来计算粒子位置,该方法允许显微镜实时跟踪单个物体。这种方法的高时空分辨率以及在测量过程中对被跟踪物体进行光学操纵的潜力有望为生物系统带来新的深入见解[4-7]。通过以标本由xy平台重新定位到显微镜中心的方式升级这种方法,具有这种长程追踪功能的粒子追踪不再局限于覆盖的视野范围。这使得能够观察活斑马鱼胚胎中线粒体的长距离运输。在这里,我们提供了用可光激活绿色荧光蛋白(mitoPAGFP)标记的线粒体的前向和逆向运动的原始数据。它与科学文章《用于研究体内脊椎动物轴突中线粒体运输的纳米分辨率实时3D轨道追踪》[8]相关。通过对轨迹应用相关分析,与均方位移方法相比,可以在偏差较小的情况下区分主动运输和暂停事件。