Morita Mitsuhiro, Susuki Jinichi, Moto Takanori, Higuchi Chitose, Kudo Yoshihisa
Laboratory of Cellular Neurobiology, School of Life Science, Tokyo University of Pharmacy and Life Science, Hachioji, Japan.
J Pharmacol Sci. 2004 Jan;94(1):25-30. doi: 10.1254/jphs.94.25.
To study calcium imaging data of cell populations that have various response patterns in peak amplitude and frequency of calcium oscillation in response to stimulation, comprehensive characterization based on statistical analysis of each response is important. In cultures of cells that are flat and in contact with each other, it is difficult to distinguish individual cells in calcium imaging data. We have developed a novel method to determine areas corresponding to individual cells in calcium imaging data. Rat neonatal cerebral astrocytes were filled with the calcium indicator Fura2, stained with acridine orange, and illuminated with UV light. The cell nuclei were clearly visualized. In addition, the images of these nuclei were useful for analyzing concentration-dependent alteration of calcium oscillation of cultured astrocytes in response to glutamate. This novel method may be useful for studying factors affecting calcium response patterns of cultured cell populations, including culture conditions, stimulus paradigms, and synthetic compounds.
为了研究细胞群体的钙成像数据，这些细胞群体在对刺激的钙振荡峰值幅度和频率方面具有各种响应模式，基于对每个响应的统计分析进行全面表征很重要。在扁平且相互接触的细胞培养物中，很难在钙成像数据中区分单个细胞。我们开发了一种新方法来确定钙成像数据中与单个细胞相对应的区域。将新生大鼠脑星形胶质细胞用钙指示剂Fura2填充，用吖啶橙染色，并用紫外光照射。细胞核清晰可见。此外，这些细胞核的图像有助于分析培养的星形胶质细胞对谷氨酸的钙振荡浓度依赖性变化。这种新方法可能有助于研究影响培养细胞群体钙反应模式的因素，包括培养条件、刺激模式和合成化合物。
