Gabriel Marion, Balle Dorothée, Bigault Stéphanie, Pornin Cyrille, Gétin Stéphane, Perraut François, Block Marc R, Chatelain François, Picollet-D'hahan Nathalie, Gidrol Xavier, Haguet Vincent
CEA, iRTSV-BGE, F-38000 Grenoble, France.
INSERM, BGE, F-38000 Grenoble, France.
Sci Rep. 2015 Oct 13;5:14532. doi: 10.1038/srep14532.
Video microscopy offers outstanding capabilities to investigate the dynamics of biological and pathological mechanisms in optimal culture conditions. Contact imaging is one of the simplest imaging architectures to digitally record images of cells due to the absence of any objective between the sample and the image sensor. However, in the framework of in-line holography, other optical components, e.g., an optical filter or a pinhole, are placed underneath the light source in order to illuminate the cells with a coherent or quasi-coherent incident light. In this study, we demonstrate that contact imaging with an incident light of both limited temporal and spatial coherences can be achieved with sufficiently high quality for most applications in cell biology, including monitoring of cell sedimentation, rolling, adhesion, spreading, proliferation, motility, death and detachment. Patterns of cells were recorded at various distances between 0 and 1000 μm from the pixel array of the image sensors. Cells in suspension, just deposited or at mitosis focalise light into photonic nanojets which can be visualised by contact imaging. Light refraction by cells significantly varies during the adhesion process, the cell cycle and among the cell population in connection with every modification in the tridimensional morphology of a cell.
视频显微镜技术具备卓越的能力,可在最佳培养条件下研究生物和病理机制的动态变化。由于在样品与图像传感器之间不存在任何物镜,接触成像成为数字记录细胞图像最简单的成像架构之一。然而,在同轴全息术的框架下,为了用相干或准相干入射光照射细胞,会在光源下方放置其他光学元件,例如滤光器或针孔。在本研究中,我们证明,对于细胞生物学中的大多数应用,包括监测细胞沉降、滚动、黏附、铺展、增殖、运动、死亡和脱离,利用具有有限时间和空间相干性的入射光进行接触成像能够达到足够高的质量。在距离图像传感器像素阵列0至1000μm的不同距离处记录细胞图案。悬浮、刚沉积或处于有丝分裂期的细胞会将光聚焦成光子纳米射流,可通过接触成像观察到。在黏附过程、细胞周期以及与细胞三维形态的每一次改变相关的细胞群体中,细胞的光折射会显著变化。