FLUORESCENCE LIFETIME IMAGING USING A CONFOCAL LASER SCANNING MICROSCOPE
FLUORESCENCE LIFETIME IMAGING USING A CONFOCAL LASER SCANNING MICROSCOPE
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DOI：
10.1002/sca.4950140305
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发表时间：
1992-05-01
期刊：
影响因子：
--
通讯作者：
LEVINE, YK
中科院分区：
文献类型：
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作者：
BUURMAN, EP;SANDERS, R;LEVINE, YK
描述了在共聚焦激光扫描显微镜中快速荧光寿命成像方法的实现。该装置采用低功率连续波（CW）氩离子激光器，配备电光斩波器，产生重复频率高达25 MHz的纳秒脉冲。时间门控检测技术可以在40 μ u-s内测量像素的寿命。给出了第一张共聚焦荧光寿命对比图。本文讨论了荧光寿命成像在多标记实验中的应用，用于区分具有重叠发射带的不同标记，探测荧光分子的局部环境以及定量荧光。
The implementation of a fast fluorescence lifetime imaging method in a confocal laser scanning microscope is described. The set up utilizes a low-power continuous wave (CW) argon ion laser equipped with an electro-optic chopper producing nanosecond pulses with a repetition rate up to 25 MHz. A time-gated detection technique enables the measurement of the lifetime of a pixel in 40-mu-s. The first confocal fluorescence lifetime contrast images are presented. Application of fluorescence lifetime imaging in multilabelling experiments for discrimination between different labels with overlapping emission bands, for probing the local environment of a fluorescent molecule, and for quantitative fluorescence are discussed.