Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX, USA.
MRC Laboratory of Molecular Biology, Cambridge, UK.
Nat Methods. 2021 Jul;18(7):829-834. doi: 10.1038/s41592-021-01175-7. Epub 2021 Jun 28.
We introduce a cost-effective and easily implementable scan unit that converts any camera-based microscope with optical sectioning capability into a multi-angle projection imaging system. Projection imaging reduces data overhead and accelerates imaging by a factor of >100, while also allowing users to readily view biological phenomena of interest from multiple perspectives on the fly. By rapidly interrogating the sample from just two perspectives, our method also enables real-time stereoscopic imaging and three-dimensional particle localization. We demonstrate projection imaging with spinning disk confocal, lattice light-sheet, multidirectional illumination light-sheet and oblique plane microscopes on specimens that range from organelles in single cells to the vasculature of a zebrafish embryo. Furthermore, we leverage our projection method to rapidly image cancer cell morphodynamics and calcium signaling in cultured neurons at rates up to 119 Hz as well as to simultaneously image orthogonal views of a beating embryonic zebrafish heart.
我们介绍了一种具有成本效益且易于实现的扫描单元，可将任何具有光学切片能力的基于摄像头的显微镜转换为多角度投影成像系统。投影成像减少了数据开销，并将成像速度提高了>100 倍，同时还允许用户即时从多个角度轻松观察感兴趣的生物现象。通过仅从两个角度快速询问样本，我们的方法还能够实现实时立体成像和三维粒子定位。我们在从细胞器到斑马鱼胚胎脉管系统的各种样本上展示了旋转盘共聚焦、晶格光片、多方向照明光片和倾斜平面显微镜的投影成像。此外，我们利用我们的投影方法以高达 119 Hz 的速率对培养神经元中的癌细胞形态动力学和钙信号进行快速成像，并同时对搏动的斑马鱼胚胎心脏的正交视图进行成像。
