Construction of a confocal microscope for real-time x-y and x-z imaging
Construction of a confocal microscope for real-time x-y and x-z imaging
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DOI：
10.1054/ceca.1999.0085
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发表时间：
1999-12-01
期刊：
影响因子：
4
通讯作者：
Parker, I
中科院分区：
文献类型：
--
作者：
Callamaras, N;Parker, I
我们描述了一个简单的“实时”激光扫描共聚焦显微镜的建设，并说明其用于快速成像的基本细胞内钙信号事件。共振扫描检流计（8 kHz）允许15或30 Hz的x-y帧采集速率，并且使用反射镜来扫描激光束允许使用真实的针孔共焦检测来提供衍射限制的空间分辨率。此外，使用压电装置来快速聚焦物镜透镜允许以类似的帧速率从厚样本获得轴向（x-z）图像。带有图像采集和图形卡的计算机将显微镜的输出转换为标准视频信号，然后可以记录在录像带上并由常规图像处理系统进行分析。该系统主要由市售部件制成，几乎不需要定制机械部件或电子电路。它的成本只有一小部分的可比商业文书，但提供更大的多功能性和类似或更好的性能。(C)哈考特出版社有限公司，1999年。
We describe the construction of a simple 'real-time' laser-scanning confocal microscope, and illustrate its use for rapid imaging of elementary intracellular calcium signaling events. A resonant scanning galvanometer (8 kHz) allows x-y frame acquisition rates of 15 or 30 Hz, and the use of mirrors to scan the laser beam permits use of true, pinhole confocal detection to provide diffraction-limited spatial resolution. Furthermore, use of a piezoelectric device to rapidly focus the objective lens allows axial (x-z) images to be obtained from thick specimens at similar frame rates. A computer with image acquisition and graphics cards converts the output from the microscope to a standard video signal, which can then be recorded on videotape and analyzed by regular image processing systems. The system is largely made from commercially available components and requires little custom construction of mechanical parts or electronic circuitry. It costs only a small fraction of that of comparable commercial instruments, yet offers greater versatility and similar or better performance. (C) Harcourt Publishers Ltd 1999.