FLUORESCENCE LIFETIME IMAGING OF INTRACELLULAR CALCIUM IN COS CELLS USING QUIN-2
FLUORESCENCE LIFETIME IMAGING OF INTRACELLULAR CALCIUM IN COS CELLS USING QUIN-2
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DOI：
10.1016/0143-4160(94)90100-7
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发表时间：
1994-01-01
期刊：
影响因子：
4
通讯作者：
NOWACZYK, K
中科院分区：
文献类型：
--
作者：
LAKOWICZ, JR;SZMACINSKI, H;NOWACZYK, K
我们描述了细胞的第一个荧光寿命图像。为了证明这种新的能力，我们基于Quin-2的Ca 2+依赖性荧光寿命测量了COS细胞中Ca 2+的细胞内图像。表观荧光寿命的测量相位调制方法，使用增益调制图像增强器和慢扫描CCD相机。我们描述的方法，以纠正在数据采集过程中的光漂白的图像，并纠正图像增强器的位置相关的响应。发现相位角Quin-2图像产生低于预期的Ca 2+浓度，这似乎是Quin-2形成荧光光产物的结果。荧光寿命成像（FLIM）不需要波长比探针，似乎为细胞的化学成像提供了新的机会。
We describe the first fluorescence lifetime images of cells. To demonstrate this new capability we measured intracellular Images of Ca2+ in COS cells based on the Ca2+-dependent fluorescence lifetime of Quin-2. Apparent fluorescence lifetimes were measured by the phase-modulation method using a gain-modulated image intensifier and a slow-scan CCD camera. We describe methods to correct the images for photobleaching during acquisition of the data, and to correct for the position-dependent response of the image intensifier. The phase angle Quin-2 images were found to yield lower than expected Ca2+ concentrations, which appears to be the result of the formation of fluorescent photoproducts by Quin-2. Fluorescence lifetime imaging (FLIM) does not require wavelength-ratiometric probes and appears to provide new opportunities for chemical imaging of cells.