Scale-free vertical tracking microscopy
Scale-free vertical tracking microscopy
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DOI：
10.1038/s41592-020-0924-7
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发表时间：
2020-08-17
期刊：
影响因子：
48
通讯作者：
Prakash, Manu
中科院分区：
文献类型：
--
作者：
Krishnamurthy, Deepak;Li, Hongquan;Prakash, Manu
与自由悬浮的生物体以及下沉颗粒相关的行为和微观过程是海洋中关键生态过程的基础。在实验室中机械地研究这种多尺度过程对显微镜提出了相当大的挑战：如何以微米级分辨率测量单细胞，同时允许它们在垂直方向上自由移动数百米？在这里，我们提出了一种无标度垂直跟踪显微镜形式的解决方案，该显微镜基于“流体动力跑步机”，沿重力轴运动不受限制。使用这种方法来弥合空间尺度，我们组装了非贴壁浮游细胞和生物体的多尺度行为数据集。此外，我们展示了“单细胞虚拟现实系统”，其中细胞行为直接控制其周围环境参数，从而实现定量行为测定。我们的方法和结果例证了一种多尺度测量的新范式，其中人们可以在微观尺度分辨率下观察和探测宏观尺度和生态相关现象。除了海洋环境之外，我们预见我们的方法将允许通过将细胞和生物体从盖玻片的限制中释放出来，对悬浮状态的细胞和生物体进行生物测量。基于“流体动力学跑步机”的无标度垂直跟踪显微镜能够以高时空分辨率测量自由悬浮生物体的长距离运动。
The behavior and microscale processes associated with freely suspended organisms, along with sinking particles underlie key ecological processes in the ocean. Mechanistically studying such multiscale processes in the laboratory presents a considerable challenge for microscopy: how to measure single cells at microscale resolution, while allowing them to freely move hundreds of meters in the vertical direction? Here we present a solution in the form of a scale-free, vertical tracking microscope, based on a 'hydrodynamic treadmill' with no bounds for motion along the axis of gravity. Using this method to bridge spatial scales, we assembled a multiscale behavioral dataset of nonadherent planktonic cells and organisms. Furthermore, we demonstrate a 'virtual-reality system for single cells', wherein cell behavior directly controls its ambient environmental parameters, enabling quantitative behavioral assays. Our method and results exemplify a new paradigm of multiscale measurement, wherein one can observe and probe macroscale and ecologically relevant phenomena at microscale resolution. Beyond the marine context, we foresee that our method will allow biological measurements of cells and organisms in a suspended state by freeing them from the confines of the coverslip.Scale-free vertical tracking microscopy based on a 'hydrodynamic treadmill' enables measuring long-range movements of freely suspended organisms with high spatiotemporal resolution.