Central Research Laboratory, Hamamatsu Photonics K.K., Hamamatsu, Shizuoka, Japan.
Genes Cells. 2021 Aug;26(8):596-610. doi: 10.1111/gtc.12876. Epub 2021 Jul 2.
Various studies have been conducted to obtain quantitative phase information based on differential interference contrast (DIC) microscopy. As one such attempt, we propose in this study a single-shot quantitative phase imaging (QPI) method by combining two developments. First, an add-on optical system to a commercialized DIC microscope was developed to perform quantitative phase gradient imaging (QPGI) with single image acquisition using a polarization camera. Second, an algorithm was formulated to reconstitute QPI from the obtained QPGI by reducing linear artifacts, which arise in simply integrated QPGI images. To demonstrate the applicability of the developed system in cell biology, the system was used to measure various cell lines and compared with fluorescence microscopy images of the same field of view. Consistent with previous studies, nucleoli and lipid droplets can be imaged by the system with greater optical path lengths (OPL). The results also implied that combining fluorescence microscopy and the developed system might be more informative for cell biology research than using these methods individually. Exploiting the single-shot performance of the developed system, time-lapse imaging was also conducted to visualize the dynamics of intracellular granules in monocyte-/macrophage-like cells. Our proposed approach may accelerate the implementation of QPI in standard biomedical laboratories.
已经有多项研究致力于通过微分干涉对比(DIC)显微镜获取定量相位信息。作为其中的一种尝试,我们在这项研究中提出了一种结合两种发展的单次定量相位成像(QPI)方法。首先,我们开发了一个附加的光学系统,将其添加到商业化的 DIC 显微镜中,以便使用偏振相机单次采集实现定量相位梯度成像(QPGI)。其次,我们制定了一种算法,通过减少简单集成的 QPGI 图像中出现的线性伪影,从获得的 QPGI 中重建 QPI。为了证明所开发系统在细胞生物学中的适用性,我们使用该系统测量了各种细胞系,并与同一视场的荧光显微镜图像进行了比较。与先前的研究一致,系统可以对核仁(nucleoli)和脂滴(lipid droplets)进行成像,其光学路径长度(OPL)更大。结果还表明,与单独使用这些方法相比,将荧光显微镜和所开发系统结合使用可能会为细胞生物学研究提供更多信息。利用所开发系统的单次拍摄性能,我们还进行了延时成像,以可视化单核细胞/巨噬细胞样细胞内颗粒的动态变化。我们提出的方法可能会加速 QPI 在标准生物医学实验室中的应用。