Construction of a two-photon microscope for video-rate Ca2+ imaging
Construction of a two-photon microscope for video-rate Ca2+ imaging
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DOI：
10.1054/ceca.2001.0246
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发表时间：
2001-12-01
期刊：
影响因子：
4
通讯作者：
Parker, I
中科院分区：
文献类型：
--
作者：
Nguyen, QT;Callamaras, N;Parker, I
我们描述了视频速率双光子激光扫描显微镜的结构，比较其性能与类似的共聚焦显微镜，并说明其用于成像局部钙离子瞬态从大脑皮层神经元切片。主要特点包括使用钛蓝宝石飞秒激光器，允许在宽（700-1000 nm）波长范围内连续调谐，共振扫描镜允许在30 Hz下采集帧，以及高效的宽场荧光检测。双光子成像在提高成像深度、减少光毒性和光漂白方面比共聚焦显微镜具有引人注目的优势，但商用仪器的高成本限制了它们的广泛采用。通过构建自己的系统，在不牺牲性能的情况下大大降低了费用，并且显微镜可以更容易地适应特定的应用。(C) 2001哈考特出版有限公司
We describe the construction of a video-rate two-photon laser scanning microscope, compare its performance to a similar confocal microscope, and illustrate its use for imaging local Ca2+ transients from cortical neurons in brain slices. Key features include the use of a Ti-sapphire femtosecond laser allowing continuous tuning over a wide (700-1000 nm) wavelength range, a resonant scanning mirror to permit frame acquisition at 30 Hz, and efficient wide-field fluorescence detection. Two-photon imaging provides compelling advantages over confocal microscopy in terms of improved imaging depth and reduced phototoxicity and photobleaching, but the high cost of commercial instruments has limited their widespread adoption. By constructing one's own system the expense is greatly reduced without sacrifice of performance, and the microscope can be more readily tailored to specific applications. (C) 2001 Harcourt Publishers Ltd.