Quantitative examination of a perfusion microscope for the study of osmotic response of cells
Quantitative examination of a perfusion microscope for the study of osmotic response of cells
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DOI：
10.1115/1.1784474
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发表时间：
2004-08-01
影响因子：
1.7
通讯作者：
Fujii, M
中科院分区：
文献类型：
--
作者：
Takamatsu, H;Komori, Y;Fujii, M
为了研究细胞的渗透反应，研制了灌注显微镜。在这个显微镜下，细胞被固定在一个安装在台上的透明腔中，并通过向腔内灌注不同浓度的溶液来暴露在各种环境中。所提供的溶液的浓度由两个变速注射泵控制，该泵提供等渗溶液和高渗溶液。在使用该系统表征细胞的渗透反应之前，使用激光干涉仪和图像处理技术定量地检测了流过腔室的NaCl溶液浓度的变化。NaCl浓度以一定的恒定速率从等渗状态突然或逐渐升高到高渗状态，从高渗状态下降到等渗状态。实验结果表明，NaCl浓度在腔室中间的横方向上几乎是均匀的，NaCl浓度的变化是可重现的。当浓度以恒定速率逐渐变化时，所测浓度的平均增减率与给定的速率相当吻合。突变速率也确定为本方法所能达到的最高极限。作为第一个使用灌注显微镜进行生物学研究的应用，我们测量了细胞暴露于高渗溶液后的体积变化。然后，通过实验与理论估算的灌注溶液中NaCl浓度变化量的体积变化量的比较，确定细胞膜的水导率。
The perfusion microscope was developed for the study of the osmotic response of cells. In this microscope, the cells are immobilized in a transparent chamber mounted on the stage and exposed to a variety of milieus by perfusing the chamber with solutions of different concentrations. The concentration of the supplied solution is controlled using two variable-speed syringe pumps, which supply an isotonic solution and a hypertonic solution. Before using this system to characterize the osmotic response of cells, the change in the concentration of NaCl solution flowing through the chamber is examined quantitatively using a laser interferometer and an image processing technique. The NaCl concentration is increased from an isotonic condition to a hypertonic condition abruptly or gradually at a given constant rate, and decreased from a hypertonic condition to an isotonic condition. It is confirmed that the concentration is nearly uniform in the cross direction at the middle of the chamber and the change in the NaCl concentration is reproducible. The average rate of increase or decrease in the measured concentration agrees fairly well with the given rate when the concentration is changed gradually at a constant rate. The rate of the abrupt change is also determined to be the highest limit achieved by the present method. As the first, application of using the perfusion microscope for biological studies, the volume change of cells after exposure to a hypertonic solution is measured. Then, the hydraulic conductivity of the cell membrane is determined from the comparison of the volume change between the experiment and the theoretical estimation for the measured change in the NaCl concentration of the perfused solution.