Department of Information and Communications, Gwangju Institute of Science and Technology, Buk-gu, Gwangju, Republic of Korea.
J Biomed Opt. 2010 Jul-Aug;15(4):045005. doi: 10.1117/1.3470104.
Monitoring a degranulation process in a live mast cell is a quite important issue in immunology and pharmacology. Because the size of a granule is normally much smaller than the resolution limit of an optical microscope system, there is no direct real-time live cell imaging technique for observing degranulation processes except for fluorescence imaging techniques. In this research, we propose optical quantitative phase microscopy (QPM) as a new observation tool to study degranulation processes in a live mast cell without any fluorescence labeling. We measure the cell volumes and the cross sectional profiles (x-z plane) of an RBL-2H3 cell and a HeLa cell, before and after they are exposed to calcium ionophore A23187 and silver nanoparticles (AgNPs). We verify that the volume and the cross sectional line profile of the RBL-2H3 cell were changed significantly when it was exposed to A23187. When 50 microg/mL of AgNP is used instead of A23187, the measurements of cell volume and cross sectional profiles indicate that RBL-2H3 cells also follow degranulation processes. Degranulation processes for these cells are verified by monitoring the increase of intracellular calcium (Ca(2+)) and histamine with fluorescent methods.
实时监测活肥大细胞脱颗粒过程在免疫学和药理学中是一个非常重要的问题。因为颗粒的大小通常比光学显微镜系统的分辨率极限小得多，除了荧光成像技术外，没有直接的实时活细胞成像技术用于观察脱颗粒过程。在这项研究中，我们提出了光学定量相位显微镜（QPM）作为一种新的观察工具，用于在没有任何荧光标记的情况下研究活肥大细胞的脱颗粒过程。我们测量了 RBL-2H3 细胞和 HeLa 细胞暴露于钙离子载体 A23187 和银纳米颗粒（AgNPs）前后的细胞体积和横截面轮廓（x-z 平面）。我们验证了当 RBL-2H3 细胞暴露于 A23187 时，其体积和横截面线轮廓发生了显著变化。当使用 50μg/mL 的 AgNP 代替 A23187 时，细胞体积和横截面轮廓的测量表明 RBL-2H3 细胞也遵循脱颗粒过程。通过用荧光方法监测细胞内钙离子(Ca(2+))和组氨酸的增加来验证这些细胞的脱颗粒过程。
