Mojiri Soheil, Isbaner Sebastian, Mühle Steffen, Jang Hongje, Bae Albert Johann, Gregor Ingo, Gholami Azam, Enderlein Jörg
III. Institute of Physics - Biophysics, Georg-August-University, 37077 Göttingen, Germany.
Max-Planck-Institute for Dynamics and Self-Organization, 37077 Göttingen, Germany.
Biomed Opt Express. 2021 May 7;12(6):3169-3180. doi: 10.1364/BOE.419099. eCollection 2021 Jun 1.
High speed volumetric optical microscopy is an important tool for observing rapid processes in living cells or for real-time tracking of sub-cellular components. However, the 3D imaging capability often comes at the price of a high technical complexity of the imaging system and/or the requirement of demanding image analysis. Here, we propose a combination of conventional phase-contrast imaging with a customized multi-plane beam-splitter for enabling simultaneous acquisition of images in eight different focal planes. Our method is technically straightforward and does not require complex post-processing image analysis. We apply our multi-plane phase-contrast microscope to the real-time observation of the fast motion of reactivated axonemes with sub-µm spatial and 4 ms temporal resolution. Our system allows us to observe not only bending but also the three-dimensional torsional dynamics of these micro-swimmers.
高速体积光学显微镜是观察活细胞中快速过程或实时跟踪亚细胞成分的重要工具。然而，三维成像能力往往以成像系统的高技术复杂性和/或苛刻的图像分析要求为代价。在这里，我们提出了一种传统相衬成像与定制多平面分束器的组合，用于在八个不同焦平面上同时采集图像。我们的方法在技术上很简单，不需要复杂的后处理图像分析。我们将多平面相衬显微镜应用于以亚微米空间分辨率和4毫秒时间分辨率实时观察重新激活的轴丝的快速运动。我们的系统不仅能让我们观察到这些微游动体的弯曲，还能观察到它们的三维扭转动力学。
