Langehanenberg Patrik, Ivanova Lyubomira, Bernhardt Ingolf, Ketelhut Steffi, Vollmer Angelika, Dirksen Dieter, Georgiev Georgi, von Bally Gert, Kemper Björn
University Medical Center of Muenster, Center for Biomedical Optics and Photonics, Robert-Koch-Strasse 45, 48149 Muenster, Germany.
J Biomed Opt. 2009 Jan-Feb;14(1):014018. doi: 10.1117/1.3080133.
Digital holographic microscopy (DHM) enables a quantitative multifocus phase contrast imaging that has been found suitable for technical inspection and quantitative live cell imaging. The combination of DHM with fast and robust autofocus algorithms enables subsequent automated focus realignment by numerical propagation of the digital holographically reconstructed object wave. In combination with a calibrated optical imaging system, the obtained propagation data quantify axial displacements of the investigated sample. The evaluation of quantitative DHM phase contrast images also enables an effective determination of lateral cell displacements. Thus, 3-D displacement data are provided. Results from investigations on sedimenting red blood cells and HT-1080 fibrosarcoma cells in a collagen tissue model demonstrate that DHM enables marker-free automated quantitative dynamic 3-D cell tracking without mechanical focus adjustment.
数字全息显微镜（DHM）能够进行定量多焦点相衬成像，已被证明适用于技术检测和定量活细胞成像。DHM与快速且强大的自动对焦算法相结合，通过数字全息重建物波的数值传播实现后续自动对焦重新对准。结合校准后的光学成像系统，所获得的传播数据可量化被研究样品的轴向位移。对定量DHM相衬图像的评估还能够有效确定细胞的横向位移。因此，可提供三维位移数据。对胶原蛋白组织模型中沉降的红细胞和HT - 1080纤维肉瘤细胞的研究结果表明，DHM能够在无需机械对焦调整的情况下实现无标记自动定量动态三维细胞跟踪。
